Phosphorylation of CLIP-170 by Plk1 and CK2 promotes timely formation of kinetochore-microtubule attachments

EMBO J. 2010 Sep 1;29(17):2953-65. doi: 10.1038/emboj.2010.174. Epub 2010 Jul 27.

Abstract

CLIP-170 is implicated in the formation of kinetochore-microtubule attachments through direct interaction with the dynein/dynactin complex. However, whether this important function of CLIP-170 is regulated by phosphorylation is unknown. Herein, we have identified polo-like kinase 1 (Plk1) and casein kinase 2 (CK2) as two kinases of CLIP-170 and mapped S195 and S1318 as their respective phosphorylation sites. We showed that a CK2 unphosphorylatable mutant lost its ability to bind to dynactin and to localize to kinetochores during prometaphase, indicating that the CK2 phosphorylation of CLIP-170 is involved in its dynactin-mediated kinetochore localization. Furthermore, we provide evidence that Plk1 phosphorylation of CLIP-170 at S195 enhances its association with CK2. Finally, we detected defects in the formation of kinetochore fibres in cells expressing the CLIP-S195A and -S1318A, but not the CLIP-S195E and -S1318D, confirming that Plk1- and CK2-associated phosphorylations of CLIP-170 are involved in the timely formation of kinetochore-microtubule attachments in mitosis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Casein Kinase II / metabolism*
  • Cell Cycle Proteins / metabolism*
  • Dynactin Complex
  • HeLa Cells
  • Humans
  • Kinetochores / metabolism*
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / metabolism*
  • Mitosis*
  • Neoplasm Proteins / metabolism*
  • Phosphorylation
  • Protein Binding
  • Protein-Serine-Threonine Kinases / metabolism*
  • Proto-Oncogene Proteins / metabolism*
  • Serine / metabolism

Substances

  • Cell Cycle Proteins
  • Dynactin Complex
  • Microtubule-Associated Proteins
  • Neoplasm Proteins
  • Proto-Oncogene Proteins
  • cytoplasmic linker protein 170
  • Serine
  • Casein Kinase II
  • Protein-Serine-Threonine Kinases
  • polo-like kinase 1