Cdc2-mediated phosphorylation of CLIP-170 is essential for its inhibition of centrosome reduplication.
CLIP-170, the founding member of microtubule "plus ends tracking" proteins, is involved in many critical microtubule-related functions, including recruitment of dynactin to the microtubule plus ends and formation of kinetochore-microtubule attachments during metaphase. Although it has been reported that CLIP-170 is a phosphoprotein, neither have individual phosphorylation sites been identified nor have the associated kinases been extensively studied. Herein, we identify Cdc2 as a kinase that phosphorylates CLIP-170. We show that Cdc2 interacts with CLIP-170 mediating its phosphorylation on Thr(287) in vivo. Significantly, expression of CLIP-170 with a threonine 287 to alanine substitution (T287A) results in its mislocalization, accumulation of Plk1 and cyclin B, and block of the G2/M transition. Finally, we found that depletion of CLIP-170 leads to centrosome reduplication and that Cdc2 phosphorylation of CLIP-170 is required for the process. These results demonstrate that Cdc2-mediated phosphorylation of CLIP-170 is essential for the normal function of this protein during cell cycle progression.
Yang
Xiaoming
X
College of Chemistry, Sichuan University, Chengdu 610064, China.
Li
Hongchang
H
Liu
X Shawn
XS
Deng
Anping
A
Liu
Xiaoqi
X
D000818:Animals; D016203:CDC2 Protein Kinase; D002453:Cell Cycle; D002460:Cell Line; D045744:Cell Line, Tumor; D018385:Centrosome / Q000648:ultrastructure*; D019926:Cyclin B / Q000737:chemistry* / Q000502:physiology*; D018844:Cyclin-Dependent Kinases; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000737:chemistry*; D008870:Microtubules / Q000378:metabolism; D009363:Neoplasm Proteins / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D010766:Phosphorylation; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D014404:Tubulin / Q000737:chemistry
D019926:Cyclin B; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D010455:Peptides; D011994:Recombinant Proteins; D014404:Tubulin; C076744:cytoplasmic linker protein 170; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; D018844:Cyclin-Dependent Kinases
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