CLIP-170 facilitates the formation of kinetochore-microtubule attachments

EMBO J. 2006 Jan 11;25(1):45-57. doi: 10.1038/sj.emboj.7600916. Epub 2005 Dec 15.

Abstract

CLIP-170 is a microtubule 'plus end tracking' protein involved in several microtubule-dependent processes in interphase. At the onset of mitosis, CLIP-170 localizes to kinetochores, but at metaphase, it is no longer detectable at kinetochores. Although RNA interference (RNAi) experiments have suggested an essential role for CLIP-170 during mitosis, the molecular function of CLIP-170 in mitosis has not yet been revealed. Here, we used a combination of high-resolution microscopy and RNAi-mediated depletion to study the function of CLIP-170 in mitosis. We found that CLIP-170 dynamically localizes to the outer most part of unattached kinetochores and to the ends of growing microtubules. In addition, we provide evidence that a pool of CLIP-170 is transported along kinetochore-microtubules by the dynein/dynactin complex. Interference with CLIP-170 expression results in defective chromosome congression and diminished kinetochore-microtubule attachments, but does not detectibly affect microtubule dynamics or kinetochore-microtubule stability. Taken together, our results indicate that CLIP-170 facilitates the formation of kinetochore-microtubule attachments, possibly through direct capture of microtubules at the kinetochore.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Chromosomes / metabolism
  • Dynactin Complex
  • Dyneins / metabolism
  • Humans
  • Kinetochores / chemistry
  • Kinetochores / metabolism*
  • Microtubule-Associated Proteins / analysis
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / chemistry
  • Microtubules / metabolism*
  • Mitosis*
  • Neoplasm Proteins / analysis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • RNA Interference
  • Spindle Apparatus / chemistry
  • Spindle Apparatus / metabolism*

Substances

  • Dynactin Complex
  • Microtubule-Associated Proteins
  • Neoplasm Proteins
  • cytoplasmic linker protein 170
  • Dyneins