Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | anti-Pericentrin (mouse monoclonal) | Abcam | Abcam Cat# ab28144, RRID:AB_2160664 | (1:500) for IF |
| Antibody | anti-Pericentrin (rabbit polyclonal) | Abcam | Abcam Cat# ab4448, RRID:AB_304461 | (1:500) for IF; (1:1000) for WB |
| Antibody | anti-CDK5RAP2 (rabbit polyclonal) | Bethyl Laboratories | Bethyl Cat# A300-554A, RRID:AB_477974 | (1:300) for IF; (1:1000) for WB |
| Antibody | anti-γ-tubulin (mouse monoclonal) | Sigma-Aldrich | Sigma-Aldrich: T6557; RRID:AB_477584 | (1:300) for IF; (1:2000) for WB |
| Antibody | anti-γ-tubulin (rabbit polyclonal) | Sigma-Aldrich | Sigma-Aldrich:T3559, RRID:AB_477575 | (1:300) for IF |
| Antibody | anti-NEDD1 (mouse monoclonal) | Abnova | Abnova Corporation Cat# H00121441-M05, RRID:AB_534956 | (1:300) for IF |
| Antibody | anti-NEDD1 (rabbit polyclonal) | Rockland | Rockland Cat# 109–401 C38S, RRID:AB_10893219 | (1:1000) for WB |
| Antibody | anti-Ninein (rabbit polyclonal) | BETHYL | Bethyl Cat# A301-504A, RRID:AB_999627 | (1:300) for IF; (1:2000) for WB |
| Antibody | anti-Ninein (mouse monoclonal) | Santa Cruz Biotechnology | Santa Cruz Biotechnology Cat# sc-376420, RRID:AB_11151570 | (1:300) for IF |
| Antibody | anti-Dynein HC (rabbit polyclonal) | Santa Cruz Biotechnology | Santa Cruz Biotechnology Cat# sc-9115, RRID:AB_2093483 | (1:300) for IF; (1:500) for WB |
| Antibody | anti-p150Glued (mouse monoclonal) | BD Biosciences | BD Biosciences Cat# 610473, RRID:AB_397845 | (1:100) for IF; (1:500) for WB |
| Antibody | anti-PCM1 (mouse monoclonal) | Santa Cruz Biotechnology | Santa Cruz Biotechnology Cat# sc-398365, RRID:AB_2827155 | (1:300) for IF |
| Antibody | anti-PCM1 (rabbit polyclonal) | Bethyl Laboratories | Bethyl Cat# A301-150A, RRID:AB_873100 | (1:300) for IF |
| Antibody | anti-AKAP450 (mouse monoclonal) | BD Biosciences | BD Biosciences Cat# 611518, RRID:AB_398978 | (1:500) for WB |
| Antibody | anti-CAMSAP2 (rabbit polyclonal) | Proteintech | Proteintech Cat# 17880–1-AP, RRID:AB_2068826 | (1:200) for IF; (1:1000) for WB |
| Antibody | anti-p53 (mouse monoclonal) | Santa Cruz Biotechnology | Santa Cruz Biotechnology Cat# sc-126, RRID:AB_628082 | (1:300) for IF; (1:1000) for WB |
| Antibody | anti-p53 (rabbit polyclonal) | BETHYL | Bethyl Cat# A300-248A, RRID:AB_263349 | (1:300) for IF |
| Antibody | anti-EB1 (mouse monoclonal) | BD Biosciences | BD Biosciences:610535; RRID:AB_397892 | (1:400) for IF |
| Antibody | anti-EB3 (rabbit polyclonal) | Martin et al., 2018; | (1:300) for IF | |
| Antibody | anti-Centrin (mouse monoclonal) | Millipore | Millipore Cat# 04–1624, RRID:AB_10563501 | (1:500) for IF |
| Antibody | anti-CEP120 (rabbit polyclonal) | Thermo Fisher Scientific | Thermo Fisher Scientific Cat# PA5-55985, RRID:AB_2639665 | (1:300) for IF |
| Antibody | anti-CEP135 (rabbit polyclonal) | Sigma-Aldrich | Sigma-Aldrich:SAB4503685; RRID:AB_10746232 | (1:300) for IF |
| Antibody | anti-CEP152 (rabbit polyclonal) | Abcam | Abcam, Cat # ab183911 | (1:300) for IF; (1:1000) for WB |
| Antibody | anti-CEP170 (mouse monoclonal) | Thermo Fisher Scientific | Thermo Fisher Scientific Cat# 41–3200, RRID:AB_2533502 | (1:200) for IF |
| Antibody | anti-CEP192 (rabbit polyclonal) | Bethyl Laboratories | Bethyl Cat# A302-324A, RRID:AB_1850234 | (1:300) for IF; (1:1000) for WB |
| Antibody | anti-GM130 (mouse monoclonal) | BD Biosciences | BD Biosciences:610823; RRID:AB_398142 | (1:300) for IF; (1:2000) for WB |
| Antibody | anti-α-tubulin YL1/2 (rat monoclonal) | Pierce | Pierce: MA1-80017; RRID:AB_2210201 | (1:300) for IF |
| Antibody | anti-α-tubulin (mouse monoclonal) | Sigma-Aldrich | Sigma-Aldrich:T5168; RRID:AB_477579 | (1:400) for IF |
| Antibody | anti-α-tubulin (rabbit monoclonal antibody) | Abcam | Abcam Cat# ab52866, RRID:AB_869989 | (1:800) for IF |
| Antibody | Anti-β-tubulin (mouse monoclonal) | Sigma-Aldrich | Sigma-Aldrich Cat# T8660, RRID:AB_477590 | (1:2000) for WB |
| Antibody | anti-CLASP1 (rabbit polyclonal) | Akhmanova et al., 2001 | (1:400) for IF | |
| Antibody | anti-CLASP2 (rabbit polyclonal) | Akhmanova et al., 2001 | (1:400) for IF | |
| Antibody | anti-CLIP-115 #2,238 (rabbit polyclonal) | Akhmanova et al., 2001 | (1:300) for IF | |
| Antibody | anti-CLIP-170 #2,360 (rabbit polyclonal) | Akhmanova et al., 2001 | (1:300) for IF | |
| Antibody | anti-ch-TOG (rabbit polyclonal) | Charrasse et al., 1998 | Dr. Lynne Cassimeris (Lehigh University, USA) |
(1:200) for IF |
| Antibody | anti-CPAP (rabbit polyclonal) | Kohlmaier et al., 2009 | Dr. Pierre Gönczy (EPFL, Switzerland) |
(1:200) for IF |
| Antibody | anti-CP110 (rabbit monoclonal) | Proteintech | Proteintech Cat# 12780–1-AP, RRID:AB_10638480 | (1:300) for IF |
| Antibody | anti-KIF1C (rabbit polyclonal) | Cytoskeleton | Cytoskeleton Cat# AKIN11-A, RRID:AB_10708792 | (1:300) for IF |
| Antibody | anti-KIF2A (rabbit polyclonal) | Ganem and Compton, 2004 | Dr. Duane Compton (Geisel School of Medicine at Dartmouth, USA) |
(1:300) for IF |
| Antibody | anti-HAUS2 (rabbit polyclonal) | Lawo et al., 2009 | Dr. Laurence Pelletier (Lunenfeld-Tanenbaum Research Institute, Canada) |
(1:200) for IF |
| Antibody | anti-BICD2 (rabbit polyclonal) | Hoogenraad et al., 2003 | (1:2500) for WB | |
| Antibody | anti-Actin (mouse monoclonal) | Millipore | Millipore Cat# MAB1501, RRID:AB_2223041 | (1:4000) for WB |
| Antibody | anti-Ku80 (mouse monoclonal) | BD Biosciences | BD Biosciences Cat# 611360, RRID:AB_398882 | (1:2000) for WB |
| Antibody | anti-LaminA/C (mouse monoclonal) | BD Biosciences | BD Biosciences Cat# 612162, RRID:AB_399533 | (1:400) for IF |
| Antibody | anti-Cytochrome C (mouse monoclonal) | BD Biosciences | BD Biosciences Cat# 556432, RRID:AB_396416 | (1:300) for IF |
| Antibody | anti-Calnexin (rabbit polyclonal) | Abcam | Abcam Cat# ab22595, RRID:AB_2069006 | (1:300) for IF |
| Antibody | Anti-Lamtor4 (rabbit monoclonal) | Cell Signaling (CST)/Bioke | Cell Signaling Technology Cat# 12284, RRID:AB_2797870 | (1:800) for IF |
| Antibody | Anti-Tom20 (mouse monoclonal) | BD Biosciences | BD Biosciences Cat# 612278, RRID:AB_399595 | (1:200) for IF |
| Antibody | IRDye 800CW/680LT secondaries | Li-Cor Biosciences | LI-COR Biosciences Cat# 926–32219, RRID:AB_1850025’ LI-COR Biosciences Cat# 926–68020, RRID:AB_10706161; LI-COR Biosciences Cat# 926–32211, RRID:AB_621843; LI-COR Biosciences Cat# 926–68021, RRID:AB_10706309 |
(1:5000) for WB |
| Antibody | Alexa Fluor 405–, 488–, and 594– secondaries | Molecular Probes/ Thermo Fisher Scientific |
Molecular Probes Cat# A-11007, RRID:AB_141374; Cat# A-11034, RRID:AB_2576217; Cat# A32723, RRID:AB_2633275; Cat# A-31553, RRID:AB_221604; Cat# A-11029, RRID:AB_138404; Cat# A-11032, RRID:AB_2534091; Cat# A-11006, RRID:AB_141373; Thermo Fisher Scientific Cat# A-11012, RRID:AB_2534079 |
(1:500) for IF |
| Sequence-based reagent | siRNA against PCNT #1 | Gavilan et al., 2018 | 5’-AAAAGCUCUGAUU UAUCAAAAGAAG-3’ |
|
| Sequence-based reagent | siRNA against PCNT #2 | Gavilan et al., 2018 | 5’-UGAUUGGACGUCA UCCAAUGAGAAA-3’ |
|
| Sequence-based reagent | siRNA against PCNT #3 | Tibelius et al., 2009 | 5’-GCAGCUGAGCUGAAGGAGA-3’ | |
| Sequence-based reagent | siRNA against CDK5RAP2 | Fong et al., 2008 | 5’-UGGAAGAUCUCCUAACUAA-3’ | |
| Sequence-based reagent | siRNA against γ-tubulin #1 | Lüders et al., 2006 | 5’-GGAGGACAUGUUCAAGGAA-3’ | |
| Sequence-based reagent | siRNA against γ-tubulin #2 | Vinopal et al., 2012 | 5’-CGCAUCUCUUUCUCAUAU-3’ | |
| Sequence-based reagent | siRNA against Ninein | Goldspink et al., 2017 | 5’-CGGUACAAUGAGUGUAGAAUU-3’ | |
| Sequence-based reagent | siRNA against PCM1 | Wang et al., 2013 | 5’-UCAGCUUCGUGAUUCUCAG-3’ | |
| Sequence-based reagent | siRNA against CEP152 | Cizmecioglu et al., 2010; Komarova et al., 2005 | 5’-GCGGAUCCA ACUGGAAAUCUA-3’ |
|
| Sequence-based reagent | siRNA against CEP120 | Ganem et al., 2005; Lin et al., 2013 | 5’-AAUAUAUCUUCU UGCAUCUCCUUCC-3’ |
|
| Sequence-based reagent | siRNA against CEP192 | Sonnen et al., 2013 | 5’-CAGAGGAAUCAAUAAUAAA –3’ | |
| Sequence-based reagent | siRNA against NEDD1 #1 | Lüders et al., 2006 | 5’-GCAGACAUGUGUCAAUUUA-3’ | |
| Sequence-based reagent | siRNA against NEDD1 #2 | Haren et al., 2006 | 5’-GGGCAAAAGCAGACAUGUG-3’ | |
| Sequence-based reagent | siRNA against DHC #1 | Splinter et al., 2010 | 5’-CGUACUCCCGUGAUUGAUG-3’ | |
| Sequence-based reagent | siRNA against DHC #2 | Splinter et al., 2010 | 5’-GCCAAAAGUUACAGACUUU-3’ | |
| Sequence-based reagent | siRNA against CAMSAP2 | Jiang et al., 2014 | 5’-GUACUGGAUAAAUAAGGUA-3’ | |
| Sequence-based reagent | siRNA against CEP170 | Stolz et al., 2015 | 5’-GAAGGAAUCCUCCAAGUCA-3’ | |
| Sequence-based reagent | siRNA against CPAP | Tang et al., 2009 | 5’-AGAAUUAGCUCGAAUAGAA-3’ | |
| Sequence-based reagent | siRNA against CLIP170 #1 | Lansbergen et al., 2004; Mimori-Kiyosue et al., 2005 | 5’-GGAGAAGCAGCAGCACAUU-3’ | |
| Sequence-based reagent | siRNA against CLIP170 #2 | Lansbergen et al., 2004; Mimori-Kiyosue et al., 2005 | 5’-UGAAGAUGUCAGGAGAUAA-3’ | |
| Sequence-based reagent | siRNA against CLIP115 #1 | Lansbergen et al., 2004 | 5’-GGCACAGCAUGAGCAGUAU-3’ | |
| Sequence-based reagent | siRNA against CLIP115 #2 | Lansbergen et al., 2004 | 5’-CUGGAAAUCCAAGCUGGAC-3’ | |
| Sequence-based reagent | siRNA against ch-TOG: | Cassimeris and Morabito, 2004; Lansbergen et al., 2004 | 5’-GAGCCCAGAGUGGUCCAAA-3’ | |
| Sequence-based reagent | siRNA against EB1 | Grigoriev et al., 2008; Lansbergen et al., 2004 | 5’-AUUCCAAGCUAAGCUAGAA-3’ | |
| Sequence-based reagent | siRNA against EB3 | Cassimeris and Morabito, 2004; Komarova et al., 2005 | 5’-CUAUGAUGGAAAGGAUUAC-3’ | |
| Sequence-based reagent | siRNA against KIF2A | Ganem et al., 2005; Grigoriev et al., 2008 | 5’-GGCAAAGAGAUUGACCUGG-3’ | |
| Sequence-based reagent | siRNA against CP110 | Cizmecioglu et al., 2010; Spektor et al., 2007 | 5'-AAGCAGCAUGAGUAUGCCAGU-3' | |
| Sequence-based reagent | siRNA against Luciferase | Lansbergen et al., 2004; Lin et al., 2013 | 5’-CGUACGCGGAAUACUUCGA-3’ | |
| Sequence-based reagent | sgRNA target CAMSAP2 | Lansbergen et al., 2004; Wu et al., 2016 | 5’-gCATGATCGATACCCTCATGA-3 | |
| Sequence-based reagent | sgRNA target p53 e2 #1 | This study | 5′-gCGTCGAGCCCCCTCTGAGTC-3′; | |
| Sequence-based reagent | sgRNA target p53 e4 #2 | This study | 5′-gCCATTGTTCAATATCGTCCG-3′; | |
| Sequence-based reagent | sgRNA target PCNT e5-1 #1 | This study | 5′-gAGACGGCATTGACGGAGCTG-3′; | |
| Sequence-based reagent | sgRNA target PCNT e5-2 #2 | This study | 5′-GCTCAACAGCCGGCGTGCCC-3′; | |
| Sequence-based reagent | p53 KO sequencing primer F | This study | 5′-TCAGACACTGGCATGGTGTT-3′; | |
| Sequence-based reagent | p53 KO sequencing primer R | This study | 5′-AGAAATGCAGGGGGATACGG-3′; | |
| Sequence-based reagent | PCNT KO sequencing primer F | This study | 5′-ATACAGCGAGGGAATTCGGG-3′; | |
| Sequence-based reagent | PCNT KO sequencing primer R | This study | 5′-TAGAATGCCCACACCGAGC-3′; | |
| Sequence-based reagent | Forward primer for PCR of tagBFP to generate pB80-FRB-TagBFP-GCN4-ppKin14 | This study | 5′-TCTCAAAGCAATTGT CGACAGGATCCGC TGGCTCCGCTGCTG GTTCTGGCGAATTCA GCGAGCTGATTA AGGAGAACA-3′; |
|
| Sequence-based reagent | Reverse primer for PCR of tagBFP to generate pB80-FRB-TagBFP-GCN4-ppKin14 | This study | 5′-ATAGCGGAGCC TGCTTTTTTGTACA CATTAAGCTTGTG CCCCAGTTTG-3′; |
|
| Sequence-based reagent | Forward primer for PCR of tagBFP to generate pB80-FRB-HA-GCN4-ppKin14 | This study | 5′-TCTCAAAGCAAT TGTCGACATACCCATA CGATGTTCCAGAT TACGCTGTGTAC AAAAAAGCAGGCTCC-3′; |
|
| Sequence-based reagent | Reverse primer for PCR of tagBFP to generate pB80-FRB-HA-GCN4-ppKin14 | This study | 5′-GGAGCCTGCTTT TTTGTACACAG CGTAATCTGG AACATCGTATGG GTATGTCGACAA TTGCTTTGAGA-3′; |
|
| Chemical compound | Centrinone B | Tocris Bioscience | Tocris Bioscience Cat # 5,690 | 125 nM |
| Chemical compound | Nocodazole | Sigma-Aldrich | Sigma-Aldrich, Cat # M1404-10MG | 10 μM |
| Chemical compound | Rapalog (A/C Heterodimerizer) | Takara | Takara, Cat # 635,056 | 50 nM (fixation), 100 nM (live imaging). |
| Chemical compound | Rapalog (B/B Homodimerizer) | Takara | Takara, Cat # 635,060 | 500 nM (fixation) |
| Chemical compound | Dynapyrazole A | Sigma-Aldrich | Sigma-Aldrich, Cat # SML2127-25MG |
5 μM |
| Chemical compound | BI2536 | Selleckchem | Selleckchem, Cat # S1109 | 500 nM |
| Chemical compound | Thymidine | Sigma-Aldrich | Sigma-Aldrich, Cat # T9250-25G | 5 mM |
| Chemical compound | proTAME | Boston Biochem | Boston Biochem, Cat # I-440 | 5 μM |
| Chemical compound | Brefeldin A | Peptrotech | Peptrotech, Cat # 2031560 | 5 µg/ml |
| Chemical compound | SiR-tubulin | Tebu-bio | Tebu-bio, Cat # SC002 | 100 nM |
| Software, algorithm | ImageJ radiality plugin |
Katrukha, 2019; https://github.com/ekatrukha/radialitymap |
|
|
| Recombinant DNA reagent | pLVX-IRES-puro (plasmid) |
Clontech | ||
| Recombinant DNA reagent | pLVX-GFP- CDK5RAP2-IRES-puro (plasmid) |
This work | ||
| Recombinant DNA reagent | pB80-FRB-TagBFP-GCN4-ppKin14 (plasmid) | This work | ||
| Recombinant DNA reagent | pB80-FRB-GFP-GCN4-ppKin14 (plasmid) | This work | ||
| Recombinant DNA reagent | pB80-FRB-HA-GCN4-ppKin14 (plasmid) | This work | ||
| Recombinant DNA reagent | 2FKBP-mCherry-CAMSAP2 (plasmid) | This work | ||
| Recombinant DNA reagent | GFP-PCNT (plasmid) | This work | ||
| Recombinant DNA reagent | 2homoFKBP-mCherry-PCNT (plasmid) | This work | ||
| Recombinant DNA reagent | GST-DmKHC(1-421)-mNeonGreen (plasmid) | This work | ||
| Cell line (Homo sapiens) | hTERT-RPE-1 | ATCC | CRL-4000 | |
| Cell line (Homo sapiens) | hTERT-RPE-1 AKAP450 knockout |
Wu et al., 2016 | ||
| Cell line (Homo sapiens) | hTERT-RPE-1 AKAP450/ CAMSAP2 knockout |
Wu et al., 2016 | ||
| Cell line (Homo sapiens) | hTERT-RPE-1 AKAP450/CAMSAP2/p53 knockout |
This work | ||
| Cell line (Homo sapiens) | hTERT-RPE-1 AKAP450/CAMSAP2/ p53/Pericentrin knockout |
This work | ||
| Cell line (Homo sapiens) | hTERT-RPE-1 AKAP450/ CAMSAP2/EB1/EB3 mutant |
This work | ||
| Cell line (Homo sapiens) | hTERT-RPE-1 AKAP450/CAMSAP2/ CDK5RAP2/MMG/ p53/Pericentrin knockout |
This work | ||
| Cell line (Homo sapiens) | HEK 293T | ATCC | CRL-11268 |