The forkhead-associated domain protein Cep170 interacts with Polo-like kinase 1 and serves as a marker for mature centrioles.
We report the characterization of Cep170, a forkhead-associated (FHA) domain protein of previously unknown function. Cep170 was identified in a yeast two-hybrid screen for interactors of Polo-like kinase 1 (Plk1). In human cells, Cep170 is constantly expressed throughout the cell cycle but phosphorylated during mitosis. It interacts with Plk1 in vivo and can be phosphorylated by Plk1 in vitro, suggesting that it is a physiological substrate of this kinase. Both overexpression and small interfering RNA (siRNA)-mediated depletion studies suggest a role for Cep170 in microtuble organization and cell morphology. Cep170 associates with centrosomes during interphase and with spindle microtubules during mitosis. As shown by immunoelectron microscopy, Cep170 associates with subdistal appendages, typical of the mature mother centriole. Thus, anti-Cep170 antibodies stain only one centriole during G1, S, and early G2, but two centrioles during late G2 phase of the cell cycle. We show that Cep170 labeling can be used to discriminate bona fide centriole overduplication from centriole amplification that results from aborted cell division.
Guarguaglini
Giulia
G
Department of Cell Biology, Max-Planck Institute for Biochemistry, D-82152, Martinsried, Germany. giulia.guarguaglini@unifoma1.it
Duncan
Peter I
PI
Stierhof
York D
YD
Holmström
Tim
T
Duensing
Stefan
S
Nigg
Erich A
EA
D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002455:Cell Division; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002502:Centrioles / Q000648:ultrastructure*; D018385:Centrosome / Q000648:ultrastructure; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D016193:G1 Phase; D016195:G2 Phase; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D047569:Laser Scanning Cytometry; D051379:Mice; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008869:Microtubule-Associated Proteins; D008938:Mitosis; D041681:NIH 3T3 Cells; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D016196:S Phase; D014162:Transfection; D020798:Two-Hybrid System Techniques; D000097869:Polo-Like Kinase 1
D018797:Cell Cycle Proteins; D016895:Culture Media, Serum-Free; D018076:DNA, Complementary; D049452:Green Fluorescent Proteins; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D000097869:Polo-Like Kinase 1; C502822:Cep170 protein, human