Distinct mechanisms govern the localisation of Drosophila CLIP-190 to unattached kinetochores and microtubule plus-ends

J Cell Sci. 2005 Aug 15;118(Pt 16):3781-90. doi: 10.1242/jcs.02504.

Abstract

CLIP-170 was the first microtubule plus-end-tracking protein to be described, and is implicated in the regulation of microtubule plus-ends and their interaction with other cellular structures. Here, we have studied the cell-cycle-dependent mechanisms which localise the sole Drosophila melanogaster homologue CLIP-190. During mitosis, CLIP-190 localises to unattached kinetochores independently of spindle-checkpoint activation. This localisation depends on the dynein-dynactin complex and Lis1 which also localise to unattached kinetochores. Further analysis revealed a hierarchical dependency between the proteins with respect to their kinetochore localisation. An inhibitor study also suggested that the motor activity of dynein is required for the removal of CLIP-190 from attached kinetochores. In addition, we found that CLIP-190 association to microtubule plus-ends is regulated during the cell cycle. Microtubule plus-end association is strong in interphase and greatly attenuated during mitosis. Another microtubule plus-end tracking protein, EB1, directly interacts with the CAP-Gly domain of CLIP-190 and is required to localise CLIP-190 at microtubule plus-ends. These results indicate distinct molecular requirements for CLIP-190 localisation to unattached kinetochores in mitosis and microtubule ends in interphase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cells, Cultured
  • Drosophila
  • Drosophila Proteins
  • Dynactin Complex
  • Dyneins / antagonists & inhibitors
  • Dyneins / metabolism
  • Genes, cdc / physiology
  • Interphase / physiology
  • Kinetochores / metabolism*
  • Microtubule-Associated Proteins / metabolism*
  • Microtubules / metabolism*
  • Mitosis / physiology*
  • Protein Binding / physiology
  • Protein Structure, Tertiary / physiology
  • Spindle Apparatus / metabolism*

Substances

  • Drosophila Proteins
  • Dynactin Complex
  • Lis-1 protein, Drosophila
  • Microtubule-Associated Proteins
  • cytoplasmic linker protein 190
  • Dyneins