The Nup107-160 complex and gamma-TuRC regulate microtubule polymerization at kinetochores

Nat Cell Biol. 2010 Feb;12(2):164-9. doi: 10.1038/ncb2016. Epub 2010 Jan 17.

Abstract

The metazoan nuclear pore complex (NPC) disassembles during mitosis, and many of its constituents distribute onto spindles and kinetochores, including the Nup107-160 sub-complex. We have found that Nup107-160 interacts with the gamma-tubulin ring complex (gamma-TuRC), an essential and conserved microtubule nucleator, and recruits gamma-TuRC to unattached kinetochores. The unattached kinetochores nucleate microtubules in a manner that is regulated by Ran GTPase; such microtubules contribute to the formation of kinetochore fibres (k-fibres), microtubule bundles connecting kinetochores to spindle poles. Our data indicate that Nup107-160 and gamma-TuRC act cooperatively to promote spindle assembly through microtubule nucleation at kinetochores: HeLa cells lacking Nup107-160 or gamma-TuRC were profoundly deficient in kinetochore-associated microtubule nucleation. Moreover, co-precipitated Nup107-160- gamma-TuRC complexes nucleated microtubule formation in assays using purified tubulin. Although Ran did not regulate microtubule nucleation by gamma-TuRC alone, Nup107-160-gamma-TuRC complexes required Ran-GTP for microtubule nucleation. Collectively, our observations show that Nup107-160 promotes spindle assembly through Ran-GTP-regulated nucleation of microtubules by gamma-TuRC at kinetochores, and reveal a relationship between nucleoporins and the microtubule cytoskeleton.

Publication types

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

MeSH terms

  • Animals
  • Cell Line
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Kinetochores / metabolism*
  • Microscopy, Fluorescence
  • Microtubules / metabolism*
  • Nuclear Pore / metabolism
  • Nuclear Pore Complex Proteins / metabolism*
  • Nuclear Proteins / metabolism*
  • Spindle Apparatus
  • Tubulin / metabolism*
  • Xenopus
  • ran GTP-Binding Protein / metabolism

Substances

  • NUP107 protein, human
  • NUP160 protein, human
  • Nuclear Pore Complex Proteins
  • Nuclear Proteins
  • Tubulin
  • ran GTP-Binding Protein