Centrosome‐Signaling Pathway Crosstalk: A Core Hub From Cellular Homeostasis to Disease

ABSTRACT

The centrosome, an evolutionarily conserved organelle in most animal cells, plays a pivotal role in fundamental processes such as cell division and ciliogenesis. Recent evidence increasingly highlights active crosstalk between the centrosome and the signaling pathways, through which cells dynamically detect and respond to diverse extracellular and intracellular cues. In this review, we summarize the roles of the centrosome in multiple signaling pathways, including Hedgehog, Wnt, and Notch that govern cellular growth, division, differentiation, and tissue homeostasis. We also explore how these interactions mold centrosomal behavior, emphasizing its function as a hub for signaling integration.

1. Centrosome Overview

1.1. The Basic Structure of the Centrosome

The centrosome is a vital organelle in animals, fungi, and several other eukaryotic lineages, serving as the major microtubule‐organizing center (MTOC) and the base of cilium during ciliogenesis (Arquint et al. 2014; Azimzadeh 2021; Jaiswal and Singh 2021; Hannaford and Rusan 2024). The centrosome comprises a pair of centrioles embedded within an amorphous proteinaceous matrix termed pericentriolar material (PCM) (Figure 1A) (Robbins et al. 1968). Over the past decades, electron microscopy and cryo‐electron tomography have elucidated the structure of the centriole as delicate and evolutionarily conserved ninefold symmetric architecture (Paintrand et al. 1992; Riparbelli et al. 2012; Guichard et al. 2010, 2012, 2013; Li et al. 2012, 2019; Greenan et al. 2018; Klena et al. 2020; Nazarov et al. 2020; Tollervey et al. 2025). Such centrioles can be categorized into canonical and noncanonical ones based on their structural and compositional characteristics (LeGuennec et al. 2021; Fu et al. 2015). In species such as Homo sapiens , Mus musculus , and Chlamydomonas reinhardtii , canonical centrioles are primarily composed of nine microtubule triplets; conversely, centrioles in Ecdysozoans—including nematodes—are characterized by short microtubule singlets or doublets, accompanied by a correlative reduction in the repertoire of centrosomal proteins (Azimzadeh and Marshall 2010; Carvalho‐Santos et al. 2011). Through proteomics and bioinformatic analyses, hundreds of centrosomal proteins have been identified (Andersen et al. 2003; Müller et al. 2010; Carden et al. 2023). Advancements in super‐resolution microscopy then have unveiled the ultrastructural architecture of these proteins, facilitating the establishment of spatiotemporal models for centrosome structure and assembly (Huang et al. 2017; Fu and Zhang 2019; Tian et al. 2021; Laporte et al. 2024). Many centrosomal proteins exhibit a concentric ring‐like distribution pattern in both human and Drosophila centrosomes, with radii increasing from the inner to outer regions (Fu and Glover 2012; Lawo et al. 2012; Mennella et al. 2012; Sonnen et al. 2012). Specifically, the Drosophila centrosome can be partitioned into five distinct zones. Zones I–IV form sequential concentric rings from the innermost to outermost regions, with Zone V localized at the distal end of the centriole (Fu and Glover 2012; Fu et al. 2015).

FIGURE 1.

Schematic structure of human centrosome and its duplication cycle. (A) Centriole microtubule walls are well organized in a nine‐fold symmetry, which are composed of nine microtubule triplets. A centriole “cap” (orange) at the distal end controls the length of the centriole and the process of ciliogenesis. Distal appendages (DA, blue) and subdistal appendages (SDA) (green) surround the distal part of the centriole, and they modulate ciliogenesis and microtubule nucleation, respectively. Pericentriolar materials (PCM, purple) locate at the proximal end of the centriole and mainly function as the platform to nucleate microtubule through the recruitment of ɣ‐TuRC and to initiate the assembly of the daughter centriole. The cartwheel structure (dark red) at the proximal end participates in the assembly of the daughter centriole. In early G1 phase, mother and daughter centrioles disengage, and a bundle of fibers (gray) links them together at their proximal end. (B) Coordinates with the cell cycle, the centrosome is strictly replicated once and segregated into the daughter cells. During early G1, a centrosome contains two centrioles which are disengaged and linked by a bundle of thin fibers. In S phase, centrosome duplication initiates; daughter centrioles assemble at the proximal end of the mother centriole and elongate through S, G2, and mitosis. In M phase, mother and daughter centrosomes separate, and PCM expands to assemble the spindle that divides chromosomes to the daughter cells. Meanwhile, daughter centrioles are converted into centrosomes that gain the ability to duplicate and recruit PCM in the next cell cycle. In the next G1, the younger mother centriole acquires the distal appendages and subdistal appendages.

Two critical complexes—distal appendages (DA) and subdistal appendages (SDA)—reside at the distal region of the mature centriole (also named mother centriole) (Paintrand et al. 1992). DA exhibits a ninefold symmetrical structure, with multiple components identified, including CEP83, CEP89, Sodium channel and clathrin linker 1 (SCLT1), Fas‐binding factor 1 (FBF1), CEP164, Ankyrin repeat domain‐containing protein 26 (ANKRD26), and other emerging candidates (Graser et al. 2007; Joo et al. 2013; Sillibourne et al. 2013; Tanos et al. 2013; Wei et al. 2013; Kanie et al. 2025). During ciliogenesis, DA plays essential roles by functioning at multiple steps, including facilitating the removal of CP110 from the mature centriole through recruitment of Tau‐tubulin kinase 2 (TTBK2), regulating ciliary‐directed vesicular transport via interactions with Rab8 and Rabin8, and mediating centriole‐to‐membrane docking (Mansour et al. 2021; Ma et al. 2023). SDA components comprise Outer dense fiber protein 2 (ODF2), CEP128, Centriolin, Coiled‐coil domain‐containing protein 68 (CCDC68), CCDC120, Ninein, and CEP170 (Chong et al. 2020). SDA anchors centriolar microtubules with one end and cytoplasmic microtubules with the other, thereby regulating centrosome positioning within the cell (Gasic et al. 2015). SDA also contributes to ciliogenesis by maintaining the interaction between Golgi apparatus and the cilia, as well as the membrane invagination surrounding the ciliary base—termed the “ciliary pocket” (Mazo et al. 2016). The distal end of centrioles harbors a “cap” that directly regulates centriole length and ciliogenesis, comprising major components such as CP110 and CEP97 (Iyer et al. 2025).

The proximal end of the mature centriole organizes the PCM (Fu and Glover 2012; Lawo et al. 2012), functioning primarily to provide a structural platform for microtubule nucleation and anchoring at the centrosome, as well as the biogenesis of new centriole (Chi et al. 2021; Dictenberg et al. 1998). Numerous PCM proteins have been identified, including pericentrin (PCNT), CEP192, CDK5 regulatory subunit‐associated protein 2 (CDK5Rap2), NEDD1 (also termed “GCP‐WD”), and γ‐tubulin (Pimenta‐Marques and Bettencourt‐Dias 2020). During interphase, the PCM exhibits a highly ordered architecture extending 200–250 nm from the centriole periphery, organized in a concentric ring‐like configuration (Fu and Glover 2012; Lawo et al. 2012; Jana 2021). Upon mitotic entry, the PCM undergoes structural transformation from “toroids” to a “matrix” structure, marked by significant expansion in volume and adoption of an amorphous arrangement (Fu and Glover 2012; Sonnen et al. 2012; Ramani et al. 2018).

Recent studies propose centriolar satellites as an integral component of the centrosome in addition to the centriole and the PCM (Tyagi et al. 2025). Centriolar satellites are spherical, electron‐dense protein granules orbiting around the centrosome, with a diameter of 70–100 nm in vertebrate cells (Sorokin 1968). Among these components, Pericentriolar material 1 (PCM1) serves as a key scaffolding molecule that orchestrates satellite assembly and structural maintenance (Begar et al. 2025). Over 600 committed and potential satellite proteins have been identified through multiple methods, including CEP290, E3 ubiquitin ligase mind bomb (MIB1), CEP131, Orofaciodigital syndrome 1 (OFD1), etc. (Gheiratmand et al. 2019; Quarantotti et al. 2019). Centriolar satellite proteins are increasingly recognized to play roles in diverse biological processes, including ciliogenesis, centrosome assembly and duplication, autophagy, microtubule organization, and cellular stress responses (Chodisetty et al. 2024; Hall et al. 2023; Joachim et al. 2017; Kodani et al. 2015; Villumsen et al. 2013).

To ensure mitotic fidelity, the centrioles must undergo precise duplication during each cell cycle, enabling daughter cells to each inherit one centrosome (Figure 1B). Centrioles are assembled from a set of evolutionarily conserved proteins, with their replication and maturation tightly regulated in a spatiotemporal manner (Fu et al. 2015; Fu and Glover 2016). During G1 phase, Polo‐like kinase 4 (PLK4) is recruited by centriolar scaffold proteins CEP192 and CEP152, forming an annular assembly platform at the proximal region of each centriole to initiate centriole duplication (Habedanck et al. 2005; Kim et al. 2013; O'Connell et al. 2001). Subsequently, SCL/TAL1 interrupting locus (STIL) localizes as a dot on the ring of PLK4 on the centriole wall (Ohta et al. 2014). PLK4 phosphorylates STIL, inducing conformational changes that enable the recruitment of Spindle assembly abnormal protein 6 (SAS6) and initiation of the cartwheel assembly (Dzhindzhev et al. 2014; Kratz et al. 2015; Lettman et al. 2013; Moyer et al. 2015). Our previous work has elucidated the molecular complexes that govern the subsequent assembly of the ninefold symmetrical centriole in Drosophila cells, identifying SAS6, CEP135, Ana1 (CEP295 in mammals), Ana3 (RTTN in mammals), Rcd4 (PPP1R35 in mammals), and Ana2 (STIL in mammals) as the core structural components (Fu and Glover 2016; Tian et al. 2021, 2022). At the distal extremity of each cartwheel spoke, STIL interacts cooperatively with Centrosomal P4.1‐associated protein (CPAP), which collaborates with CEP120 to regulate centriole microtubule elongation and control the centriole length (Comartin et al. 2013; Lin et al. 2013). Upon completion of the duplication cycle, the fibrous networks connecting the two mature centrioles dissolve, initiating centrosome separation. This process is crucially mediated by NIMA‐related kinase 2 (Nek2), which phosphorylates Centrosomal Nek2‐associated protein 1 (C‐Nap1) to disrupt centrosome cohesion (Fry et al. 1998; Mayor et al. 2002). Centrosome tethering is also maintained by additional structural proteins, including β‐catenin, Rootletin, CEP68, CEP215, Conductin/Axin2, Leucine‐rich repeat‐containing protein 45 (LRRC45), and centlein (Prigent and Uzbekov 2022). Following separation, the two independent centrosomes each comprise a nascent daughter centriole and an orthogonal mother centriole, precisely positioned in perpendicular orientation (Prigent and Uzbekov 2022). This is mediated by CDK5Rap2, which forms direct interaction networks with CEP68 and PCNT to stabilize centriole engagement (Pagan et al. 2015). During prometaphase, CEP68 undergoes ubiquitin‐proteasomal degradation mediated by the β‐TrCP and Polo‐like kinase 1 (PLK1) kinase activity (Pagan et al. 2015); upon mitotic exit, separase activation triggers proteolytic cleavage of PCNT (Ito et al. 2025; Lee and Rhee 2012). These lead to CDK5Rap2 dissociation from the centrosome and the completion of centriole disengagement (Pagan et al. 2015). In late interphase and mitosis, CEP135, CEP295 (Ana1 in Drosophila), and CEP152 (Asl in Drosophila) sequentially decorate daughter centrioles, forming a molecular network (Fu et al. 2016; Izquierdo et al. 2014; Wang et al. 2011). Through CEP152 recruiting PLK4, this molecularly nascent centriole is transformed into a replication‐competent mother centriole, accomplishing centriole‐to‐centrosome conversion (Fu and Glover 2016; Fu et al. 2016).

To summarize, recent studies have extensively elucidated the intricate organization and structure of the centrosome. However, novel centrosomal proteins continue to be discovered and characterized, and their functions and conservation across organisms await further investigation (Gupta et al. 2015). Meanwhile, although the ninefold symmetry of the centrosome is highly conserved across species, variations exist among different model organisms. For instance, somatic cells in Drosophila contain centrioles with doublet microtubules (A‐ and B‐tubules) connected by A‐A linkers; whereas in humans and some other organisms, centrioles consist of triplet microtubules (A‐, B‐, and C‐tubules) connected via A‐C linkers (Greenan et al. 2018). The mechanistic basis of these organizational differences and their biological significance thus warrant deeper exploration.

1.2. Classical Functions of the Centrosome

1.2.1. The Centrosome Coordinates the Cell Cycle

The centrosome assembles spindle pole during mitosis and mediates spindle assembly, a process in which the PCM plays a pivotal role by functioning as the anchoring site for the minus ends of microtubules (Kollman et al. 2011). The microtubule nucleation activity of the PCM is mediated by the γ‐tubulin ring complex (γ‐TuRC), whose core building block is the γ‐tubulin small complex (γ‐TuSC). In mammals, the γ‐TuSC comprises γ‐tubulin complex protein 2 (GCP2), γ‐tubulin complex protein 3 (GCP3), and two γ‐tubulin molecules (Murphy et al. 1998). As a constitutive component of the PCM, γ‐tubulin is robustly recruited to the PCM during mitosis, where it assembles into γ‐TuRCs that facilitate centrosomal microtubule nucleation (Khodjakov and Rieder 1999). This recruitment relies on the kinase activities of PLK1 and Aurora kinase A (Aurora A) (Ohta et al. 2021). Numerous centrosomal proteins mediate γ‐TuRC recruitment, including PCNT, A‐kinase anchor protein 9 (AKAP9), CEP192, GCP‐WD, and CDK5Rap2 (Petry and Vale 2015). Concurrently, the centrosome recruits spindle pole components Nuclear mitotic apparatus protein (NuMA) and Targeting protein for Xklp2 (TPX2) to facilitate bipolar spindle formation, thereby ensuring the equitable segregation of chromosomes between daughter cells (Garrett et al. 2002; Merdes et al. 1996). Additionally, the centrosome regulates cell cycle progression by serving as a docking platform for cell cycle regulatory factors such as Cyclin‐dependent kinase 1 (CDK1), Cyclin B, and PLK1 (Doxsey et al. 2005).

Studies in Drosophila germline stem cells (GSCs), neuroblasts, and mouse neural progenitors (NPs) have revealed that the centrosome executes critical functions in asymmetric cell division (ACD) (Gonzalez 2021). The maintenance of GSC stemness in Drosophila depends on their attachment to somatic hub cells and the functional divergence between the old and young centrosomes within the same cell (also referred to as mother and daughter centrosomes). By generating a mitotic spindle oriented perpendicular to the hub, GSCs undergo asymmetric division, yielding one daughter cell that remains attached to the hub (retaining stem cell identity) and another that dissociates and differentiates into a gonialblast (GB). The differential behavior of mother and daughter centrosomes dictates the divergent fates of these daughter cells: during G1 phase, the centrosome localizes near the hub interface; in G2 phase, following duplication and disengagement, the mother centrosome maintains its position near the hub, while the daughter migrates to the opposite side of the cell; upon cytokinesis, the mother centrosome is inherited by the self‐renewing GSC, whereas the daughter centrosome is passed to the differentiating GB (Yamashita et al. 2003, 2007). Meanwhile, investigations into Drosophila neural stem cells also highlight age‐dependent functional differences between old and young centrosomes in controlling spindle orientation. Mediated by Partner of inscuteable (Pins), the young centrosome organizes an aster and remains near the apical cortex early in the cell cycle, while the old centrosome loses PCM and microtubule‐organizing capacity until it migrates to the basal cortex. Subsequently, the apical centrosome is retained in the stem cell, and the basal one is allocated to the differentiating daughter cell (Rebollo et al. 2007; Januschke et al. 2011). In the ventricular zone (VZ) of the developing mouse neocortex, radial glia progenitors undergo ACD to produce self‐renewing radial glia (remaining in the VZ) and differentiating cells (subsequently migrating out of the VZ). This process is regulated by centrosomal asymmetry: progenitors preferentially inherit the old centrosome, while the new centrosome is allocated to the differentiating daughter cell. The accurate execution of this division process depends on the presence of centrosomal protein Ninein (Wang et al. 2009).

1.2.2. The Centrosome Serves as Foundation for Cilium Assembly

Cilia are present in the majority of vertebrate cell types. These antenna‐like sensory organelles, localized at the cell surface, function in signal perception and transduction (Lacigová and Čajánek 2025). Cilia can be categorized into motile cilia and primary cilia. Motile cilia mediate mechanical functions such as propulsion (e.g., sperm flagella), whereas primary cilia predominantly serve as signaling hubs for multiple pathways (Anvarian et al. 2019; Lee and Ostrowski 2021). Ciliogenesis initiates during the G0 or G1 phase of the cell cycle, during which the centrosome—also termed the “basal body”—serves as the structural foundation for ciliary assembly (Vertii et al. 2016). Primary cilia exhibit an evolutionarily conserved architecture, with a core composed of microtubule doublets arranged in ninefold symmetry at the proximal region, extending from the microtubule triplets of the basal body (Mill et al. 2023). Key centrosomal components, including DA, SDA, distal “cap,” and centriolar satellite, play critical roles in ciliogenesis. Ciliary abnormalities lead to multiple disorders including Bardet‐Biedl syndrome (BBS), Joubert syndrome, and Meckel syndrome, collectively termed “ciliopathies” (Bettencourt‐Dias et al. 2011).

Beyond its well‐known roles as the primary MTOC regulating cell division and as the basal body supporting cilium growth, the centrosome also possesses additional functions. These include actin nucleation, regulation of cell polarity, participation in neurogenesis, and involvement in phagocytosis, though the specific mechanisms underlying these roles remain to be further elucidated (Bornens 2021; Stötzel and Kiermaier 2022). Additionally, numerous proteins involved in diverse intracellular processes localize to and around the centrosome, modulating the transduction of associated signaling pathways (Uzbekov and Avidor‐Reiss 2020). In this review, we systematically summarize the interactions between the centrosome (and its associated proteins) and multiple signaling pathways (e.g., Hedgehog, Wnt and Notch). We propose that the centrosome is critically integrated with diverse cellular processes, serving as a signaling hub within the cell.

1.3. Centrosome‐Associated Diseases

The proper spatiotemporal biogenesis and maintenance of the centrosome are critical for preserving cellular, tissue, organ, and even organismal homeostasis. Dysregulation in centrosome number, structure, or function can trigger the development of various diseases. Altered centrosome numbers, for instance, have been documented in a wide range of cancers, including breast, ovarian, and prostate cancers, etc. (Hsu et al. 2005; Lingle et al. 1998; Wang et al. 2020). This association has also been validated in multiple animal models, including Drosophila and Mus musculus (Basto et al. 2008; Levine et al. 2017). Abnormalities in centrosome numbers can disrupt chromosome segregation during mitosis, thereby contributing to chromosomal instability (CIN), aneuploidy, tumorigenesis, and metastasis (Kiermaier et al. 2024). Additionally, impairments in the structure and function of the centrosome can disrupt ciliogenesis, thereby contributing to a range of developmental disorders (Farcy et al. 2023; Chen et al. 2021). Mutations in centrosomal proteins such as CPAP, WD repeat‐containing protein 62 (WDR62), CDK5Rap2, and Ribosomal RNA‐processing protein 7 homolog A (RRP7A) can lead to microcephaly that causes cerebral and neural developmental disorders (Gabriel et al. 2016; Zhang et al. 2019; Bond et al. 2005; Farooq et al. 2020).

Despite these insights into the link between centrosomal dysregulation and disease, many critical questions remain unresolved. Do cells with abnormal centrosomes employ long‐lasting mechanisms to promote tumorigenesis? What are the long‐term consequences of aberrant activation of related signaling pathways? How can recent research findings be translated into therapies for centrosome‐related diseases? Importantly, could centrosomes also be linked to other diseases? Relevant evidence has shown that various viruses accumulate at centrosomes during their entry into the cell nucleus, with retroviruses even completing assembly there (Naghavi and Walsh 2017). The centrosome functions as a site for aggresome accumulation—structures involved in protein turnover, autophagy, and host defense—and some viruses exploit this feature to facilitate their replication (Wileman 2007). These observations highlight the centrosome's role in host‐pathogen interactions, raising the possibility of its involvement in immune‐related pathological processes.

2. Hedgehog Signaling Pathway

2.1. Overview of Hedgehog Signaling Pathway

The Hedgehog (Hh) signaling pathway is an evolutionarily conserved intercellular communication system that plays critical regulatory roles in animal development, tissue homeostasis regulation, and organ regeneration (Goodrich and Scott 1998; Bertrand and Dahmane 2006; Dessaud et al. 2008; Jing et al. 2023; Singh et al. 2015). It carries out these functions by modulating specific transcriptional programs that govern cellular proliferation and differentiation (Roberts et al. 2017). Disruption of the Hh signaling pathway is linked to numerous human disorders, including congenital abnormalities such as holoprosencephaly and Pallister‐Hall syndrome, as well as various cancers (Jiang 2022).

The Hh gene was initially identified in Drosophila melanogaster . Its mutation induces segment polarity defects and ectopic bristle formation in larvae, resulting in a spiked embryonic morphology resembling a hedgehog—hence the name “hedgehog” (Niisslein‐Volhard and Wieschaus 1980). In Drosophila, a single Hh gene orchestrates developmental patterning, whereas mammals have evolved three paralogous Hh genes: Sonic (SHH), Indian (IHH), and Desert (DHH), each with distinct expression patterns and functional specializations (Bangs and Anderson 2017). SHH is critically involved in nervous system development (Douceau et al. 2023); IHH predominantly regulates skeletal development (Shimoyama et al. 2007); and DHH seems to be restricted to the gonads (Bitgood et al. 1996).

Notably, both SHH and IHH signaling depend functionally on the centrosome and primary cilia to activate downstream cascades. In the absence of Hh ligands, the transmembrane receptor Patched (PTCH) constitutively suppresses Smoothened (SMO) activity by inhibiting its translocation to the ciliary membrane, thereby maintaining pathway quiescence (Rohatgi et al. 2007). Downstream transcription factor Gli3 undergoes sequential phosphorylation by a kinase cascade involving cAMP‐dependent kinase (PKA), Casein kinase 1 (CK1) and Glycogen synthase kinase‐3β (GSK‐3β) (Chen et al. 1998; Jia et al. 2002; Price and Kalderon 2002). This post‐translational modification triggers Gli3's partial proteasomal degradation mediated by the SCF E3 ubiquitin ligase complex, ultimately generating a truncated GLI repressor form (GLI‐R) that suppresses transcriptional activation of target genes (Tempé et al. 2006). Upon Hh ligand binding to PTCH, SMO inhibition is relieved, enabling SMO translocation to the ciliary membrane and initiation of downstream signaling (Rohatgi et al. 2007). Consequently, Gli2 is released from Suppressor of fused homolog (SUFU), allowing its conversion to the activator form (GLI‐A) and subsequent nuclear translocation (Tukachinsky et al. 2010). GLI‐A then initiates transcription of target genes such as Cyclin D, Cyclin E, and Myc, thereby mediating critical developmental processes including cellular proliferation and differentiation (Figure 2) (Duman‐Scheel et al. 2002; Jiang 2022). Previous studies have established that Hh signaling critically depends on primary cilia for proper function (Bangs and Anderson 2017; Kopinke et al. 2021; Zhang and Beachy 2023); this aspect will not be elaborated further in the current context. Instead, we emphasize that Hh signal transduction is also mechanistically linked to the centrosome.

FIGURE 2.

Vertebrate centrosome regulates Hedgehog signaling pathway. The centrosome is also termed “basal body” during ciliogenesis, on which grows transition zone and axoneme. The “Y‐link” and transition fibers (distal appendages) provide a semi‐isolated environment for cilia to ensure precise signal transduction. When the Hedgehog (Hh) pathway is off, PTCH locates on the ciliary membrane and inhibits SMO. SUFU constitutively binds to Gli3, and the complex is together transported to the basal body. Ciliary and centrosomal PKA phosphorylates Gli3, resulting in the partial degradation of Gli3 (mediated by centrosomal SCF complex) and converts it to the transcription repressor form (GLI‐R). Gli2 is restricted in the centrosomal site to prevent its activation and translocation into the nucleus. Upon binding with the Hh ligand, the Hh pathway is switched on, and PTCH exits from the cilia, allowing SMO to accumulate in the cilia. Gli2 is dissociated from SUFU and transported to the basal body, where PKA is inactivated. Gli2 is activated (termed “GLI‐A”) and released from the centrosome; subsequently, it enters the nucleus to turn on the transcription of Hh target genes such as Myc, Cyclin D, and Cyclin E.

2.2. Centrosomes Are Profoundly Involved in Hedgehog Signal Transduction

2.2.1. Direct Functions: Centrosomes Directly Regulate Hedgehog Signaling Pathway

PKA, a critical signaling component in the Hh pathway, is intimately associated with the centrosome (Figure 2). It is a heterotetramer composed of two catalytic subunits (PKA‐C) and two regulatory subunits (PKA‐R). Upon cAMP binding to PKA‐R, PKA‐C dissociates from PKA‐R and becomes activated (Taylor et al. 2022). The released PKA‐C phosphorylates Gli3, triggering its proteolytic processing into the GLI repressor (GLI‐R), thereby maintaining the Hh signaling pathway in a quiescent state (Turnham and Scott 2016). Different PKA subunits exhibit distinct subcellular localization patterns. The RI subunit of PKA (PKA‐RI) is located in primary cilia (Mick et al. 2015), while the RII subunit (PKA‐RII) is specifically positioned at centrosomes mediated by AKAP9 (AKAP450) (Barzi et al. 2010; Terrin et al. 2012). The centrosome‐specific localization of PKA‐C indicates that the phosphorylation of GLI likely occurs predominantly at the centrosome (Barzi et al. 2010), either before ciliary entry or after export from cilia. However, considering the enrichment of adenylyl cyclases (enzymes that produce cAMP) within cilia (Vuolo et al. 2015) and the ciliary localization of PKA‐RI mediated by Gpr161 (Bachmann et al. 2016), PKA may translocate into cilia by sensing local cAMP concentrations, subsequently releasing PKA‐C to phosphorylate Gli3 in cilia. Previous studies have demonstrated a significant reduction in GLI‐R levels upon cilia‐targeted inhibition of PKA activity, demonstrating that Gli3 phosphorylation may also partially occur within the ciliary compartment (Mick et al. 2015). Regardless, the centrosome provides an enriched platform for PKA, ensuring efficient phosphorylation of Gli3 proteins either within cilia or at the centrosome. This post‐translational modification drives the conversion of Gli3 to GLI‐R, thereby preventing transcriptional activation of downstream genes and maintaining the Hh signaling pathway in its quiescent state.

The partial degradation of phosphorylated Gli3 is also associated with the centrosome (Figure 2). This proteolytic processing relies on the SCF complex‐mediated ubiquitination pathway. The three major components of the SCF complex are S‐phase kinase‐associated protein 1 (SKP1), Cullin 1 (CUL1), and F‐box protein (Zeng et al. 2025). CUL1 functions as a scaffold protein that connects other components within the complex. SKP1 interacts with CUL1 and the F‐box protein. β‐TrCP serves as the F‐box protein responsible for the partial proteolysis of Gli3 (Tempé et al. 2006). Notably, SKP1, CUL1, and β‐TrCP are localized to the centrosome (Freed et al. 1999; Rogers et al. 2009). Additionally, multiple proteasomal components (e.g., 20S proteasome core particles, PA700, PA28, and ubiquitin) are enriched in the pericentrosomal region, suggesting that the partial degradation of Gli3 may occur at centrosomal sites (Fabunmi et al. 2000). Collectively, it is proposed that the spatial co‐localization of PKA and SCF complexes at the centrosome enhances the efficiency of GLI‐to‐GLI‐R conversion.

Moreover, the centrosome also plays a critical role in preventing abnormal activation of Gli2. A minor fraction of Gli2 undergoes proteolytic processing via the same mechanism that mediates Gli3 degradation (Pan et al. 2006). In the absence of PKA, Gli2 accumulates at the cilium tip (Tuson et al. 2011), implying that the translocation of Gli2 to the nucleus may be partially blocked. Cep164 maintains the centrosomal localization of Gli2 in the absence of Hh ligand through direct binding. Upon Hh pathway activation, Gli2 dissociates from the centrosome and translocates to the nucleus to activate the transcription of downstream target genes (Fushimi et al. 2023).

In conclusion, the centrosome acts as an anchoring platform, providing docking sites for molecules that convert Gli3 to GLI‐R and prevent Gli2 activation. This dual regulatory mechanism further prevents aberrant activation of target gene transcription under Hh signal‐deficient conditions, ensuring precise spatiotemporal control of signal transduction. Further investigations may optimize antibody sensitivity and develop innovative methodologies to reinvestigate the putative ciliary localization of PKA‐C. Mass spectrometry combined with co‐immunoprecipitation could be applied to identify potential ciliary interaction partners of PKA‐C. Furthermore, the development of phosphorylation‐specific Gli3 antibodies will enable mapping of spatiotemporal modification dynamics, while interaction proteomics will facilitate characterization of the centrosome‐associated degradation machinery.

2.2.2. Indirect Functions: Centrosomes Orchestrate Hedgehog Signaling Pathway via Ciliogenesis and Formation of Semi‐Compartmental Environment

The centrosome orchestrates cilia development, thereby providing the structural basis for Hh signal transduction. Ciliogenesis progresses through the following steps. Initially, distal appendage vesicles (DAVs) anchor to the DA and subsequently fuse to form the ciliary vesicle (CV). The CP110‐CEP97 complex is then removed from the centrosome. Intraflagellar transport (IFT) proteins are recruited to the basal body to mediate the transport of materials essential for ciliary assembly. Finally, the simultaneous elongation of axonemal microtubules and ciliary membrane completes cilia formation (Lacigová and Čajánek 2025). Certain centrosomal proteins can modulate ciliogenesis and consequently affect Hh signaling without inducing significant structural or functional perturbations to the centrosome. Below, we summarize key centrosomal proteins influencing ciliogenesis and Hh signaling, which involve DA, SDA, distal “cap”, and satellite proteins (Tables 1 and 3).

TABLE 1.

Functions of centrosomal components in ciliogenesis.

Protein	Localization	Functions in ciliogenesis	References
CEP83/CCDC41	DA	Recruit ciliary vesicles; anchor basal body to membrane; accumulate Rabin8 at centrosome	Tanos et al. (2013); Cuenca et al. (2019)
CEP164	DA	Recruit TTBK2	Rosa E Silva et al. (2022)
Chibby	DA	Promote ciliary vesicle formation; dock basal body to membrane	Burke et al. (2014)
DZIP1	DA	Promote ciliogenesis; recruit IFT machinery to DA	Wang, Low, et al. (2013)
FBF1	DA	Accumulate Rabin8 at centrosome	Cuenca et al. (2019)
LRRC45	DA	Establish transition zone; promote docking of Rab8 GTPase‐positive vesicles to centrosome	Kurtulmus et al. (2018)
SCLT1	DA	Promote ciliogenesis	Lee et al. (2022)
ANKRD26	DA	Promote ciliogenesis	Kanie et al. (2025)
CEP89/CEP123	DA; SDA	Promote ciliary vesicle formation	Sillibourne et al. (2013)
ODF2	DA; SDA	Remove CP110; enable transition fiber formation	Ishikawa et al. (2005); Tateishi et al. (2013); Otto and Hoyer‐Fender (2023)
CEP170	SDA	Interact with dynein‐2 complex; promote ciliogenesis	Weijman et al. (2024)
Kif2a	SDA	Regulate the length of cilia	Zhang et al. (2019)
Kif3a	SDA	Participate in basal foot formation	Kodani et al. (2013)
Ndel1	SDA	Regulate the length of cilia; negatively regulate ciliogenesis	Inaba et al. (2016)
CEP97	distal “cap”	Negatively regulate ciliogenesis	Spektor et al. (2007)
CEP162	distal “cap”	Promote transition zone assembly	Wang, Tay, et al. (2013)
CP110	distal “cap”	Negatively regulate ciliogenesis	Xie et al. (2024)
CPAP	distal “cap”	Regulate the length of cilia	Wu and Tang (2012)
C2cd3	distal “cap”; satellite	Dock ciliary vesicles to centrosome; recruit multiple essential ciliogenic proteins	Ye et al. (2014)
PCM1	satellite	Promote departure of CP110 and CEP97 from centrosome; trigger ciliogenesis	Hall et al. (2023)
CEP290	satellite	Promote ciliary vesicle maturation	Kobayashi et al. (2014)
OFD1	satellite	Promote ciliogenesis	Ferrante et al. (2006)
CCDC66	satellite	Promote ciliogenesis; recruit BBS4; regulate axonemal length	Conkar et al. (2017); Odabasi et al. (2023)

TABLE 3.

Centrosome‐associated components affecting signaling pathways.

Protein	Pathway involved	Functions	References
CEP164	Hedgehog signaling pathway	Control Gli2 activity	Fushimi et al. (2023)
CCDC66	Hedgehog signaling pathway	Regulate hedgehog pathway activation	Odabasi et al. (2023)
DZIP1	Hedgehog signaling pathway	Promote cytoplasmic retention of transcription factor Gli3	Wang, Low, et al. (2013); Wang, Tay, et al. (2013)
Aurora A	Wnt signaling pathway PI3K/Akt signaling pathway	Interact with Axin Phosphorylate Akt	Subramaniyan et al. (2016); Katsha et al. (2015)
Nuf	Notch signaling pathway	Bind with Rab11	Emery et al. (2005)
Numb	Notch signaling pathway	Enable Notch binding activity	Louie et al. (2004) Hadjihannas et al. (2010)
Rab11	Notch signaling pathway	Form stable recycling endosomes to recycle Delta	Wang et al. (2014)
CRMP	Notch signaling pathway	Enrich Rab11 around nucleus to recycle Delta	Thompson et al. (2022); Ligon et al. (2001)
MIB1	Notch signaling pathway	Regulate the trafficking of Notch ligands	Tozer et al. (2017)
TACC3	Notch signaling pathway	Inhibit nuclear translocation of NICD	Bargo et al. (2010)

Meanwhile, the centrosome establishes a semi‐compartmental environment for cilia, thereby maintaining the proper localization of Hh signaling molecules within cilia or the cytoplasm. The transition zone (referring to the region between basal body and axoneme) contains “Y‐links” and DA (termed “transition fibers” during ciliogenesis) that separate the cilia from the cytoplasm (Anderson 1972; O'Toole et al. 2003; Tateishi et al. 2013; Tony Yang et al. 2015; Zhang et al. 2024). Nine transition fibers originate from basal body microtubules, connecting to the ciliary membrane with approximately 60 nm spacing between adjacent fibers (Garcia‐Gonzalo and Reiter 2017). These structures restrict diffusion of membrane proteins and large soluble proteins between the cytoplasm and the cilia (Nachury et al. 2010). This selective permeability creates a semi‐isolated compartment in cilia, providing the structural basis for the selective transportation of Hh pathway components between the cytoplasm and the cilia. The BBSome is a complex of BBS proteins that functions as a transporter for membrane proteins between the ciliary and plasma membranes (Jin et al. 2010). While the function of IFT is crucial for cilia formation (Pigino 2021), the BBSome is generally nonessential for ciliogenesis, suggesting its role in transporting a specific subset of transmembrane proteins to the cilia (Jin et al. 2010). Previous studies have demonstrated that Hh signaling molecules PTCH1, SMO, and Gpr161 are transported across the transition zone via BBSome‐mediated mechanisms and subsequently exported from the cilia, ensuring accurate subcellular localization of these components (Seo et al. 2011; Yee et al. 2015).

In summary, the centrosome provides the structural basis for ciliogenesis and physically separates the cytoplasm from the ciliary matrix through “Y‐links” and transition fibers, thereby establishing a barrier for precise subcellular localization of Hh signaling molecules. Current studies have investigated the roles of centrosomal proteins in ciliogenesis and their effects on Hh pathway activation or a few Hh pathway components, but the molecular mechanisms underlying the direct associations between centrosomal proteins and Hh pathway components remain poorly characterized. Moreover, although previous research has partially elucidated interactions between the BBSome and Hh pathway molecules, the mechanisms governing their bidirectional transport across the ciliary gate remain unclear. Future studies could focus on the biophysical properties of the transition zone to dissect the mechanisms underlying its semi‐permeability. Additionally, immunofluorescence, transcriptomic, and proteomic analyses could be applied to systematically examine how centrosomal proteins regulate the spatiotemporal dynamics of distinct Hh signaling components.

2.3. Hedgehog Signaling Molecules Affect Centrosome Activity

While the centrosome plays pivotal roles in Hh signaling transduction, Hh signaling components in turn regulate centrosome activity. Recent research has demonstrated that SUFU, a critical component of the Hh signaling pathway, can operate independently of the canonical Hh signaling pathway. It prevents centrosome overduplication through promoting CDK2‐dependent phosphorylation of CP110 (Zhuang et al. 2021). In chick neuroepithelial cells, SHH signaling and its downstream effectors maintain symmetric PKA distribution at the centrosome to ensure symmetric proliferative cell divisions. This spatial regulation preserves progenitor identity in daughter cells. Furthermore, SHH signaling upregulates centrosomal proteins (e.g., PCNT) to facilitate centrosome maturation (Saade et al. 2017). Overall, current research on the regulation of the centrosome by Hh signaling pathway components remains limited, primarily focusing on the effects of specific molecules rather than systematic investigation. Future studies should broaden the scope to comprehensively assess the impacts of Hh pathway components on centrosome activity.

3. Wnt Signaling Pathway

3.1. Overview of Wnt Signaling Pathway

The Wnt signaling pathway is a highly conserved regulatory pathway that controls embryonic development, tissue homeostasis, and cell proliferation (Liu et al. 2022). It can be divided into canonical and non‐canonical pathways. The canonical pathway, also known as the Wnt/β‐catenin pathway, has its core in the regulation of β‐catenin stability. In the absence of Wnt ligands, β‐catenin is phosphorylated by the “destruction complex” composed of Adenomatous polyposis coli protein (APC), Axin, GSK‐3β, and CK1, and is subsequently targeted for proteasomal degradation (Aberle et al. 1997; Peifer and Polakis 2000). The phosphorylation of β‐catenin is initiated by CK1, which mediates phosphorylation at the Ser45 residue of β‐catenin. Next, GSK‐3β further phosphorylates β‐catenin at the Ser33, Ser37, and Thr41 residues (MacDonald et al. 2009; Nusse and Clevers 2017). Finally, the phosphorylated serine 33 and 37 residues of β‐catenin are recognized by the E3 ubiquitin ligase β‐TrCP, leading to ubiquitination and subsequent proteasomal degradation of β‐catenin (Hart et al. 1999; Liu et al. 2002). When the extracellular Wnt ligands (Wnt proteins) bind to the membrane receptor Frizzled and its co‐receptor, Low‐density lipoprotein receptor related protein 6 (LRP6), the Wnt signaling pathway is activated (Liu et al. 2022). This interaction prompts the assembly of a Wnt‐Frizzled‐LRP6 complex, which recruits the cytoplasmic scaffolding protein Dishevelled (DVL). Subsequent phosphorylation and activation of LRP6 ensue, facilitating translocation of the Axin destruction complex to the plasma membrane (MacDonald et al. 2009; Liu et al. 2022). The β‐catenin is then not degraded and enters the nucleus, where it drives the transcription of target genes via TCF/LEF transcription factors (Behrens et al. 1996; Korinek et al. 1998). The noncanonical pathways rely on cytoskeletal reorganization and/or calcium signaling to regulate cell migration (Veeman et al. 2003). They are best known for the Wnt/planar cell polarity (PCP) pathway discovered in Drosophila, which, for example, controls the planar cell polarity in the Drosophila wing epithelium through core Frizzled/PCP factors influencing the cytoskeleton (Koca et al. 2022). Recent studies have revealed that the centrosome interacts with the Wnt signaling pathway through various mechanisms, playing a critical role in the spatiotemporal dynamics of β‐catenin, the efficiency of signal transduction, and the pathogenesis of associated diseases.

3.2. Localization of Wnt Signaling Pathway Components in Centrosomes

β‐catenin serves as a central component of the canonical Wnt signaling pathway, coordinating bipolar spindle assembly during mitosis by using the centrosome as a structural platform (Mbom et al. 2013). Initial studies have demonstrated that the overexpression of β‐catenin leads to centrosomal dysfunction and the loss of microtubule cortical anchorage (Ligon et al. 2001). During interphase, phosphorylated β‐catenin primarily localizes to the mother centrosome (marked by ε‐tubulin), while during the M phase, it is recruited to the daughter centrosome (Huang et al. 2007). Cellular localization studies have revealed that β‐catenin interacts with Rootletin and C‐Nap1 during interphase, maintaining centrosome connectivity (Bahmanyar et al. 2008; Kaplan et al. 2004). In C. elegans , SYS‐1 (β‐catenin homolog) undergoes microtubule‐dependent transport orchestrated by Dynein, which potentiates rapid centrosomal enrichment and subsequent degradation of SYS‐1 by the centrosomal proteasome (Thompson et al. 2022). Additionally, Dynein can bind to β‐catenin and may tether microtubules at adherens junctions (Ligon et al. 2001). Furthermore, β‐catenin recruits γ‐tubulin to the centrosome, forming a complex with Axin1 that is involved in microtubule nucleation at the centrosome (Fumoto et al. 2009; Mbom et al. 2013).

The centrosome facilitates the degradation of β‐catenin by recruiting and nucleating the destruction complex, thereby inducing local phosphorylation and ubiquitination of β‐catenin, which accelerates its proteasome‐dependent degradation (Lach et al. 2022; Vora et al. 2020). APC, the protein scaffold of the destruction complex, is predominantly localized to the mother centrosome (Louie et al. 2004), where it forms a complex with Axin2 to cooperatively regulate β‐catenin (Hadjihannas et al. 2010). Proteomic investigations and proximity‐dependent biotinylation (BioID) have demonstrated hundreds of functionally significant interactions between centrosome proteins and Wnt signaling effectors (Gupta et al. 2015; Jakobsen et al. 2011). These interactions have been systematically cataloged in prior reviews (Bryja et al. 2017), and will not be discussed here.

In summary, it can be observed that phosphorylated β‐catenin at the centrosome promotes centrosome maturation while simultaneously undergoing self‐degradation. These two processes may appear contradictory on the surface. Subtle temporal differences likely exist between these distinct behaviors. Additionally, the differential timing of localization on the mother and daughter centrosomes may influence the functional outcomes of β‐catenin. Future studies could utilize time‐lapse fluorescence microscopy with photoconvertible tags to quantify the kinetics of β‐catenin recruitment and degradation on mother versus daughter centrosomes across consecutive cell cycles. Additionally, engineered centrosome‐targeting peptides fused to β‐catenin degradation domains could selectively disrupt its localization timing on specific centrosome populations, enabling functional dissection of spatiotemporal effects on centrosome maturation. These possibilities require further investigation to elucidate the underlying mechanisms.

3.3. Wnt Signaling Pathway Participates in Centrosome Separation

During centrosome maturation, the separation of mother and daughter centrosomes is critical for proper cell division. β‐catenin is involved in centrosome separation, with its regulatory function primarily mediated by a phosphorylation‐dependent network. During interphase, emerging evidence suggests that GSK‐3β and Axin2 phosphorylate β‐catenin at the Ser33/Ser34/Thr41 sites on the centrosome, thereby stabilizing β‐catenin at the centrosome rather than targeting it for ubiquitin‐proteasome degradation (Chilov et al. 2011; Hadjihannas et al. 2010). During G2/M phase, CDK1 phosphorylates both PLK1 and ODF2. Phosphorylated ODF2 serves as a docking site for PLK1 recruitment and loses its inhibitory function, which reduces centrosomal β‐catenin accumulation during interphase (Yang et al. 2018). Nek2 is activated by phosphorylated PLK1 (Faragher and Fry 2003) and phosphorylates specific residues (Ser33/Ser37/Thr41) on β‐catenin. This blocks the GSK‐3β‐mediated ubiquitination and degradation of β‐catenin and stabilizes its accumulation at the centrosome (Bahmanyar et al. 2008; Mbom et al. 2014). Nek2 also phosphorylates C‐Nap1 and Rootletin, triggering their dissociation from the centrosome. Upon Rootletin dissociation, β‐catenin bound to Rootletin is released and can bind to a centrosomal site independently of Rootletin (Bahmanyar et al. 2008). Phosphorylated β‐catenin binds to the centrosome rather than the region between centrosomes, thereby promoting centrosome separation during mitosis (Bahmanyar et al. 2008; Yang et al. 2018). In the same phase, Axin2 undergoes proteasomal degradation (Bryja et al. 2017), accelerating centrosome separation.

Upon activation of the Wnt signaling pathway, the stability of β‐catenin is enhanced, its centrosomal localization is promoted, and it accelerates centrosome separation via the Nek2‐β‐catenin‐PLK1 axis, thereby facilitating efficient mitosis (Mbom et al. 2013). Simultaneously, Aurora A interacts with Axin, inhibiting the destruction complex and resulting in increased nuclear accumulation of β‐catenin (Subramaniyan et al. 2016). In the absence of Wnt signaling, centrosomes facilitate the formation of localized “degradation platforms” by recruiting destuction complex, which promotes the phosphorylation and proteasome‐dependent degradation of β‐catenin (Lach et al. 2022).

During cell division, centrosomes coordinate spindle pole polarity and signaling pathway activity by dynamically recruiting and releasing signaling molecules. The DVL localizes to the centrosome through its DIX domain and regulates the activity of Nek2 (Cervenka et al. 2016). At the G2/M phase, DVL is phosphorylated by Nek2 and released into the cytoplasm. The phosphorylated DVL binds to C‐Nap1, assisting in the release of C‐Nap1 from the centrosome (Cervenka et al. 2016). In addition to phosphorylation modifications centered on Nek2 and the destruction complex, centrosome‐localized β‐catenin can also be phosphorylated by Spleen tyrosine kinase (Syk) at Tyr142. This modification promotes the centrosomal localization of β‐catenin and inhibits its nuclear translocation (Bhardwaj et al. 2018). Aurora A exerts regulatory control over GSK‐3β via phosphorylation of Akt, thereby modulating β‐catenin activity through this signaling cascade (Katsha et al. 2015). Steroidogenic factor 1 (SF‐1, NR5A1) can maintain centrosome stability by inhibiting the activity of DNA‐dependent protein kinase (DNA‐PK), which prevents the accumulation of β‐catenin at the centrosome (Wang et al. 2014). Upon SF‐1 depletion, activated DNA‐PK/Akt signaling results in enhanced GSK‐3β phosphorylation, followed by β‐catenin accumulation at the centrosome, ultimately leading to centrosome separation (Wang et al. 2014).

The Wnt pathway critically contributes to centrosome separation processes by modulating the stability of β‐catenin, which is essential for Wnt‐mediated cellular proliferation (Figure 3). The Nek2‐β‐catenin‐PLK1 axis constitutes a critical regulatory component during bipolar spindle assembly, influencing the segregation of mother and daughter centrosomes. However, the physical mechanism by which the distinct forms of β‐catenin mediate centrosome separation through dynamic binding and dissociation with Rootletin and C‐Nap1 remains to be elucidated. Furthermore, contradictory findings persist regarding how site‐specific phosphorylation transitions at distinct β‐catenin residues modulate its stability. Moreover, there is insufficient evidence to demonstrate how centrosomes affect the establishment and transduction of canonical Wnt signaling, though they can recruit the degradation complex to degrade β‐catenin and impact the asymmetric inheritance of β‐catenin in daughter cells of C. elegans (Vora and Phillips 2015). Additionally, centrosome‐derived cilia can influence Wnt signal transduction but are not essential for activating the core Wnt/β‐catenin pathway (Ocbina et al. 2009). Notably, whether additional interacting factors are involved in the regulatory process of β‐catenin and the intricate relationship network among these distinct interaction partners remains unclear. Current investigations predominantly rely on antibody‐based fluorescence assays, whereas future studies should prioritize biochemical and structural approaches to elucidate the precise mechanisms governing β‐catenin‐mediated centriole separation. Future studies could also employ proteomic approaches to conduct comprehensive investigations into these interacting factors and their interconnections, thereby elucidating the complexity of β‐catenin regulatory mechanisms.

FIGURE 3.

Wnt signaling pathway participates in centrosome separation. During interphase, centrioles are connected by a filamentous network composed of C‐Nap1 and Rootletin. Axin2 is localized to the centrosome via binding to C‐Nap1 and forms a complex with APC and GSK‐3β that phosphorylates β‐catenin to maintain centrosomal cohesion. ODF2 interacts with Axin1, enhancing the stability of the destruction complex and reducing centrosomal β‐catenin accumulation. Nek2 and PLK1 kinase activities are suppressed to prevent premature centrosome separation. DVL slowly accumulates at the centrosome alongside other centrosomal linker proteins. Axin1 and APC additionally contribute to microtubule dynamics. During G2/M phase, CDK1 phosphorylates PLK1 and ODF2. Phosphorylated ODF2 serves as a docking site for PLK1 recruitment and loses its inhibitory function. PLK1 subsequently phosphorylates Nek2 and Axin1. Nek2 phosphorylates β‐catenin, C‐Nap1, and Rootletin, inducing disassembly of the filamentous network. DVL is also phosphorylated by Nek2, dissociates from the centrosome, and facilitates C‐Nap1 release from the centrosome. Concurrently, Axin2 undergoes proteasomal degradation, leading to disassembly of the destruction complex. Upon Wnt pathway activation, Aurora A interacts with Axin, modulating the assembly of the β‐catenin destruction complex. Arrows: activation; bars: inhibition.

3.4. Dysregulation of Centrosome‐Wnt Signaling Crosstalk in Tumorigenesis

Abnormal centrosome dynamics, such as the formation of multiple centrosomes, are closely associated with Wnt signaling dysfunction in tumor progression. β‐catenin deficiency leads to centrosome number abnormalities and chromosomal segregation errors in embryonic stem (ES) cells (Raggioli et al. 2014), while the aberrant accumulation of DVL disrupts Nek2 kinase activity, inducing monopolar spindle defects (Cervenka et al. 2016). Furthermore, cancer‐associated mutations, such as the Akt‐insensitive mutations of the Daple protein, disrupt the compartmentalized distribution of β‐catenin between centrosomes and adherens junctions, thereby promoting sustained Wnt signaling activation and tumor invasion (Aznar et al. 2017; Luo et al. 2019). Mutations in Wnt signaling components represent a prevalent mechanism contributing to oncogenesis, and numerous therapeutic approaches targeting this pathway have been developed. Notably, centrosomal abnormalities mediated by mutations in Wnt pathway‐associated proteins may emerge as a novel therapeutic target in cancer, given the critical coordination between β‐catenin‐mediated centrosome functions and cell cycle progression.

4. Notch Signaling Pathway

4.1. Overview of Notch Signaling Pathway

The Notch signaling pathway is an evolutionarily conserved cell–cell communication system that regulates cell fate determination through receptor‐ligand interactions (Kopan and Ilagan 2009). The Notch gene, first discovered in Drosophila melanogaster with notched wings, encodes a transmembrane receptor at regions of contact between adjacent cells and is activated by ligands (such as Delta/Jagged family) (Artavanis‐Tsakonas et al. 1983; Metz and Bridges 1917). Receptor activation requires two sequential proteolytic cleavages: first, an extracellular cleavage at the S2 site by the ADAM metalloproteinase family, followed by a subsequent intramembranous cleavage at the S3 site by γ‐secretase (De Strooper et al. 1999; Zolkiewska 2008). These cleavages release the Notch intracellular domain (NICD), which translocates into the nucleus (Schroeter et al. 1998). The NICD binds to the CSL transcription factor (CBF1/RBP‐Jκ). This complex recruits the co‐activator Mastermind‐like protein 1 (MAML1) and initiates transcription of target genes belonging to the Hes/Hey family (Kopan and Ilagan 2009; Sasai et al. 1992). Accumulating evidence shows that Notch signaling exhibits dual context‐dependent roles in carcinogenesis; its dysregulation drives malignancy by promoting epithelial‐mesenchymal transition, angiogenesis, metastasis, chemoresistance, and cancer stemness, while modulating metabolic reprogramming and tumor microenvironment equilibrium (Shi et al. 2024). It is worth noting that the temporal and spatial specificity of Notch signaling is determined not only by ligand‐receptor interactions but also by intricate orchestration at the organelle level. The centrosome orchestrates processes such as cell division polarity, vesicle trafficking, and kinase localization, thereby contributing to the regulation of Notch pathway activity.

4.2. Regulation of Notch Signaling by Centrosome‐Mediated Cell Polarity

The centrosome directly affects the spatial distribution of Notch signaling by orchestrating the establishment of cell polarity, thereby dictating the lineage commitment of cellular fate. In the process of asymmetric cell division of Drosophila sensory organ precursor (SOP) cells, the Notch signaling pathway mediates critical events in cell fate determination (Figure 4) (Furman and Bukharina 2011). Within the dorsal thoracic region, SOPs undergo oriented mitosis along the Drosophila body axis, yielding posterior pIIa cell and anterior pIIb daughter cell (Fichelson and Gho 2003; Gho et al. 1999). These secondary precursors then differentiate through terminal divisions into specialized cell types, including the external and internal sensory organ cells (Fichelson and Gho 2003; Gho et al. 1999). Nuf, a Rab11‐binding protein, is critical for the formation of cycling endosomes. During the asymmetric division of the SOP cell, the centrosome recruits Nuf near the anterior cortex, where Numb is also recruited, establishing an asymmetric distribution (Betschinger et al. 2003; Emery et al. 2005). Rab11 maintains uniform cellular distribution throughout division. After division, Nuf and Numb localize to the daughter cell that later becomes PIIb cell, derived from the anterior cortex. In the pIIb cell, Rab11 enriches around centrosomes to form stable recycling endosomes near the nucleus through Collapsin response mediator protein (CRMP) (Emery et al. 2005; Jauffred et al. 2013). Conversely, the other daughter cell, which becomes the PIIa cell, lacks Nuf and thus cannot form recycling endosomes (Emery et al. 2005). The Delta ligand is delivered to the PIIb cell interface via Rab11‐mediated endocytic recycling, activating the Notch pathway in pIIa daughter cells and inhibiting it in pIIb cells, thereby promoting the specification of pIIa cell fate (Emery et al. 2005). MIB1 is a ligase that regulates the trafficking of Notch ligands. In chick neural progenitor cells, MIB1 interacts with PCM1 and Azelaic acid induced 1 (AZI1) (Tozer et al. 2017). During mitosis, PCM1 and AZI1 maintain an asymmetric distribution, whereas MIB1 achieves symmetric localization via a Golgi‐derived supplemental pool (Tozer et al. 2017). In asymmetric divisions of neural progenitors, MIB1 compensatory mechanisms are abrogated, resulting in asymmetric partitioning inherited by nascent neurons. These MIB1‐positive neurons subsequently mediate Notch trans‐activation, maintaining sibling cells in an undifferentiated state (Tozer et al. 2017).

FIGURE 4.

Model of centrosome‐mediated Notch signaling during asymmetric cell division in Drosophila sensory organ precursor cells. During asymmetric division of the Drosophila sensory organ precursor (SOP) cell, the centrosome recruits Nuf near the anterior cortex, establishing asymmetric distribution, while Numb is recruited to the anterior cortex. Rab11 maintains a uniform cellular distribution throughout division. Following division, Nuf and Numb localize to the PIIb cell, where Rab11‐enriched centrosomes will form stable recycling endosomes via CRMP. In contrast, the PIIa cell lacks Nuf and exhibits no Rab11 accumulation, resulting in the asymmetric distribution of Rab11 between the two daughter cells. The PIIb cell activates Delta activity and regulates Delta trafficking, thereby establishing asymmetric Notch signaling. Delta on the PIIb cell interacts with Notch on the PIIa cell, activating Notch signaling in the PIIa cell to determine cell fate.

The centrosome plays a crucial role in asymmetric division by mediating the asymmetric distribution of Notch signaling. However, distinct mechanisms likely exist in different cell types during unequal division and differentiation, and additional regulatory factors require identification. Moreover, current research on the regulation of centrosomal activity by Notch signaling remains limited. Further investigation is warranted into how Notch signaling influences centrosomes in differentiated daughter cells following asymmetric distribution.

4.3. Centrosome‐Resident Proteins Participate in Notch Signaling Pathway

Several centrosome‐resident proteins have been reported to be directly involved in the activation and inhibition of the Notch receptor. Human Nek6 (hNek6) associates with the centrosome via its N‐terminal domain and potentially modulates the Notch signaling pathway through interactions with Centromere‐binding protein 1 interacting corepressor (CIR) and SNW domain containing 1 (SNW1) (Vaz Meirelles et al. 2010). In addition, Transforming acidic coiled‐coil protein‐3 (TACC3) can inhibit the nuclear translocation of NICD by binding to the Notch4/Int3 CDC10/Ankyrin repeats (Bargo et al. 2010). However, current evidence lacks direct demonstration of physical interactions between centrosomal components and Notch‐associated proteins. Future investigations employing proteomic approaches could elucidate the potential functional linkages between centrosome and Notch protein localization and activity.

5. Other Pathways Related to Centrosome

5.1. Ca²⁺‐Mediated Cellular Signaling Pathway

Calcium ions (Ca²⁺), the ubiquitous secondary messengers in cellular systems, play critical roles in regulating diverse biological processes, including cell proliferation and differentiation (Ulengin‐Talkish and Cyert 2023). Ca²⁺ flux dysregulation constitutes a central pathological mechanism in cardiac and vascular disorders, such as impaired contractility and fatal cardiac arrhythmias (Marks 2013). Within centrosomal compartments, calcium‐binding proteins, including calmodulin and centrin, are essential for mitotic progression. Biochemical studies have demonstrated spatiotemporal dynamics of centrosomal Ca²⁺ during mitotic progression (Helassa et al. 2019). Two calcium‐binding proteins, calmodulin (CaM) and centrin, have been reported to be essential for mitotic progression (Ulengin‐Talkish and Cyert 2023). Calcineurin (CN), a Ca²⁺/calmodulin‐dependent protein phosphatase, requires coordinated binding of Ca²⁺ and CaM for activation. POC5, an integral component of the protein scaffold maintaining structural integrity within the centrosomal lumen, has been shown to directly interact with CN through its conserved PxIxIT motif. Importantly, CN enzymatic activity significantly influences POC5 subcellular distribution within the centrosomal compartment (Tsekitsidou et al. 2023). While in vitro studies have demonstrated CN‐mediated dephosphorylation of POC5, in vivo validation of this regulatory mechanism remains to be fully established (Tsekitsidou et al. 2023). Additionally, how Ca²⁺‐binding proteins regulate centrosomal activity via Ca²⁺ and how centrosomes influence cellular Ca²⁺ signal transmission or gradient establishment remains unstudied and warrants in‐depth exploration.

5.2. PI3K/Akt Signaling Pathway

The PI3K/Akt (also known as Protein kinase B, PKB) signaling pathway plays critical roles in fundamental cellular processes including growth and survival (Kalous et al. 2023). It represents one of the most frequently activated signaling pathways in human cancers, with multifactorial activation mechanisms commonly linked to chemoresistance (Glaviano et al. 2023). As a key serine/threonine protein kinase in this cascade, Akt plays pivotal roles in mitosis. It localizes to spindle poles, participates in spindle assembly checkpoint control, modulates centrosomal integrity and composition, and contributes to cytokinesis regulation (Kalous et al. 2023). During the G2/M transition, insulin‐like growth factor signaling activates PI3‐kinase (Andjelkovic et al. 1997), culminating in Akt phosphorylation and subsequent activation (Shtivelman et al. 2002). This catalytic cascade induces phosphorylation‐mediated inactivation of GSK‐3β, thereby promoting centrosome separation. Notably, Akt also maintains centrosomal integrity in oocytes, suggesting conserved centrosomal functionality in meiotic resumption (Kalous et al. 2006). In Drosophila embryos, Akt orchestrates centrosome migration, mitotic spindle orientation, and proper spindle morphogenesis (Kalous et al. 2023). Mechanistically, Akt mediates phosphorylation‐dependent inactivation of Zw3 (GSK‐3β homolog) within the embryonic cortical region, thereby sustaining adequate levels of Armadillo (β‐catenin homolog) and APC2 (Buttrick et al. 2008). This regulatory axis stabilizes interactions between End‐binding protein 1 (EB1) and microtubules, facilitating dynein‐dependent final stages of centrosome separation and precise spindle alignment (Buttrick et al. 2008).

5.3. Nuclear Factor κB (NF‐κB) Signaling Pathway

Nuclear factor κB (NF‐κB), a pivotal transcription factor, orchestrates signaling cascades indispensable for maintaining cellular homeostasis and modulating immune responses (Hayden and Ghosh 2008). The canonical NF‐κB pathway is implicated in diverse disease contexts, with comprehensive reviews detailing its roles across pathological conditions (Schnappauf and Aksentijevich 2020; Yu et al. 2020). Experimental evidence demonstrates that NF‐κB governs the expression of genes required for centrosome duplication, with PLK4 being identified as a direct transcriptional target (Ledoux et al. 2013). CEP55, a microtubule‐bundling protein essential for mitotic progression, exhibits pronounced overexpression in multiple solid malignancies, where it functionally promotes neoplastic cell proliferation and invasive potential (Peng et al. 2017). Notably, the NF‐κB/IκBα signaling axis is found to be activated in CEP55‐transduced pancreatic cancer (PANC) cells, whereas its activity is suppressed upon CEP55 silencing (Peng et al. 2017).

5.4. MAP Kinase (MAPK) Signaling Pathway

The MAP kinase (MAPK) pathway, also termed the RAF–MEK–ERK signaling cascade, constitutes an evolutionarily conserved signaling network central to the regulation of cell proliferation (Ullah et al. 2022). Dysregulation of this pathway, including overexpression and aberrant activation of its receptors, is frequently observed in colorectal cancer (Fang and Richardson 2005). Functionally, the pathway operates via a conserved tripartite kinase module. Signaling is initiated by activation of receptor tyrosine kinases (RTKs) and RAS, culminating in MAPK phosphorylation. Extracellular signal‐regulated kinases (ERKs), a subfamily of MAPKs, are subject to stringent spatiotemporal regulation, with Raf‐1 activation serving as a critical regulatory node in this process (Dhillon and Kolch 2002). Raf kinase inhibitory protein (RKIP or PEBP), a well‐characterized suppressor of the Raf/MEK/MAP kinase cascade, has recently been shown to associate with centrosomes and kinetochores in mammalian cells, where it modulates spindle assembly checkpoint (SAC) signaling (Eves et al. 2007). Within the androgen receptor–Src signaling module, androgen stimulation triggers MEK/ERK pathway activation, which in turn promotes centrosomal accumulation of γ‐tubulin, enhances microtubule nucleation, and facilitates mitotic spindle organization (Colello et al. 2012).

In conclusion, while relatively few studies have addressed how the above pathways are connected with centrosomes, emerging evidence indicates that centrosomes and their associated proteins are capable of influencing the signaling cascades. The precise mechanistic underpinnings remain incompletely defined, underscoring the need for additional experimental validation. Centrosomes, as dynamic platforms for protein assembly and biochemical reactions, facilitate intercompartmental signal transduction within cells, thereby integrating and propagating signals across distinct cellular domains. Future investigations leveraging multi‐omics approaches may shed light on both the functional roles of centrosomes within signaling networks and how their dynamic activity modulates pathway architecture.

6. Conclusions and Perspectives

The centrosome, the pivotal organelle in mitotic division and ciliogenesis of animal cells, plays an indispensable role in cellular proliferation and cell polarity. Its dynamic behavior is subjected to rigorous regulatory mechanisms. Our review focuses on canonical cellular signaling pathways to elucidate the dual regulatory relationship between the centrosomes and the signaling cascades—specifically, how centrosomes influence pathway activities and conversely how signaling pathways modulate centrosomal functions (Tables 2 and 3). Our summary analysis reveals that across nearly all investigated pathways, the centrosome seems to be a molecular hub facilitating interactions with pathway components or associated proteins.

TABLE 2.

Components of signaling pathways regulating centrosome activity.

Protein	Pathway involved	Functions	References
SUFU	Hedgehog signaling pathway	Prevent centrosome overduplication through promoting CDK2‐dependent phosphorylation of CP110	Zhuang et al. (2021)
β‐catenin	Wnt signaling pathway	Control centrosome separation; recruit γ‐tubulin to centrosome	Ligon et al. (2001); Fumoto et al. (2009); Mbom et al. (2013)
APC	Wnt signaling pathway	Scaffold of the degradation complex; form a complex with Axin2	Louie et al. (2004) Hadjihannas et al. (2010)
Axin1	Wnt signaling pathway	Involve in microtubule nucleation	Ledoux et al. (2013)
Axin2	Wnt signaling pathway	Phosphorylate β‐catenin at ‐S33/S34/T41; inhibit centrosome separation	Eves et al. (2007)
GSK‐3β	Wnt signaling pathway	Phosphorylate β‐catenin at S33/S34/T41; inhibit centrosome separation	Chilov et al. (2011); Hadjihannas et al. (2010)
CK1	Wnt signaling pathway	Phosphorylate β‐catenin at Ser45	Nusse and Clevers (2017)
DVL	Wnt signaling pathway	Bind with C‐Nap1 to assist ‐its release from‐ centrosome	Cervenka et al. (2016)
CN	Ca2+ mediated cellular signaling pathway	Interact‐ with POC5	Tsekitsidou et al. (2023)
Akt	PI3K/Akt signaling pathway	Modulate centrosome integrity and composition	Kalous et al. (2023)
NF‐κB	NF‐κB signaling pathway	Govern the expression of PLK4	Ledoux et al. (2013)
RKIP	MAPK signaling pathway	Modulate SAC signaling	Eves et al. (2007)

Current research on the interplay between centrosome and signaling pathways remains largely unidirectional, predominantly focusing on either centrosomal regulation of signaling or pathway‐mediated centrosomal modulation. For instance, whether Notch signaling reciprocally influences centrosomal dynamics remains unexplored. We propose that this regulatory relationship constitutes a bidirectional, dynamic network requiring comprehensive investigation. However, inherent complexities arise from both the structural sophistication of centrosomes and the vast molecular networks comprising individual pathways. Present observations primarily document partial component‐centrosome interactions, leaving more intricate regulatory networks awaiting elucidation. Critical mechanistic questions persist unresolved, including the precise functional implications of distinct β‐catenin phosphorylation sites at centrosomes and the specific aspects of centrosome maturation mediated by SUFU. Current methodologies predominantly rely on fluorescence staining techniques, lacking molecular biochemical validation and genetic evidence, necessitating cautious interpretation of existing findings. Advanced gene‐editing platforms, when paired with specialized library screening, will enable systematic interrogation of how signaling molecules sculpt centrosome dynamics and how centrosomes modulate signaling output as signaling hubs. Super‐resolution microscopy, integrated with next‐generation live‐cell imaging, will further refine real‐time visualization of spatiotemporal interplay between centrosomal proteins and signaling effectors, clarifying how pathways coordinate centrosome behavior during cell cycle progression and cell fate specification. Multi‐omics integration will continue to map centrosome‐associated signaling networks with greater precision, illuminating how post‐translational modifications and protein–protein interactions bridge centrosomal structure to signaling cascades. Meanwhile, AI‐driven protein structure prediction, coupled with functional validation, will refine models of key molecular interfaces, including those between centrosomal kinases and their signaling substrates. Looking ahead, future efforts will focus on defining how centrosomes integrate diverse signaling inputs to maintain cellular homeostasis, and how their dysregulation drives pathogenesis in diseases like cancer and ciliopathies. These insights will deepen understanding of centrosomes as central nodes in signaling networks.

Author Contributions

M.P. and J.L. conducted the research and wrote the manuscript. M.P. prepared the figures. J.F. provided guidance on research design and manuscript organization.

Conflicts of Interest

The authors declare no conflicts of interest.

Pan, M. , Li J., and Fu J.. 2026. “Centrosome‐Signaling Pathway Crosstalk: A Core Hub From Cellular Homeostasis to Disease.” Cytoskeleton 83, no. 8: 578–600. 10.1002/cm.70027.

Data Availability Statement

Data sharing not applicable to this article as no datasets were generated or analysed during the current study.