GCP-WD is a gamma-tubulin targeting factor required for centrosomal and chromatin-mediated microtubule nucleation

Nat Cell Biol. 2006 Feb;8(2):137-47. doi: 10.1038/ncb1349. Epub 2005 Dec 25.

Abstract

The gamma-tubulin ring complex (gammaTuRC) is a large multi-protein complex that is required for microtubule nucleation from the centrosome. Here, we show that the GCP-WD protein (originally named NEDD1) is the orthologue of the Drosophila Dgrip71WD protein, and is a subunit of the human gammaTuRC. GCP-WD has the properties of an attachment factor for the gammaTuRC: depletion or inhibition of GCP-WD results in loss of the gammaTuRC from the centrosome, abolishing centrosomal microtubule nucleation, although the gammaTuRC is intact and able to bind to microtubules. GCP-WD depletion also blocks mitotic chromatin-mediated microtubule nucleation, resulting in failure of spindle assembly. Mitotic phosphorylation of GCP-WD is required for association of gamma-tubulin with the spindle, separately from association with the centrosome. Our results indicate that GCP-WD broadly mediates targeting of the gammaTuRC to sites of microtubule nucleation and to the mitotic spindle, which is essential for spindle formation.

Publication types

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

MeSH terms

  • Antigens / metabolism
  • Cell Line
  • Centrosome / metabolism*
  • Chromatin / metabolism*
  • HeLa Cells
  • Humans
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / drug effects
  • Microtubules / metabolism*
  • Mitosis / physiology
  • Mutation / genetics
  • Nocodazole / pharmacology
  • Phosphorylation
  • Protein Binding
  • RNA Interference
  • RNA, Small Interfering / genetics
  • Spindle Apparatus / metabolism
  • Transfection
  • Tubulin / genetics
  • Tubulin / metabolism*

Substances

  • Antigens
  • Chromatin
  • Microtubule-Associated Proteins
  • NEDD1 protein, human
  • RNA, Small Interfering
  • TUBGCP2 protein, human
  • Tubulin
  • pericentrin
  • Nocodazole