Figure 4.

NACA dephosphorylation leads to recruitment of novel interactants. A, Venn diagram showing overlap and comparison between the sets of NACA-associated proteins identified by MS. Network analysis and visualization of the identified NACA interactants based on curated protein– interaction databases (GeNets). Interactants unique to PP1A overexpression are delineated within the red rectangle. Closely connected protein clusters are color coded (green, intermediate filaments; purple, cajal bodies; orange, cytoskeleton; blue, mRNA processing; red, transcriptional regulation; interactants that are not part of defined clusters are shown in gray). B, table of the PP1A-dependent NACA interactome. NACA-associated proteins identified by MS in the presence or absence of PPP1CA. C, validation of NACA interaction with BTF3 in a PP1A-dependent manner. HEK293T cells expressing NACA-FLAG and/or MYC-PPP1CA were lysed and immunoprecipitated (IP) with anti-FLAG antibody beads and probed for endogenous BTF3. Overexpression of MYC-PPP1CA induces interaction of NACA and BTF3.