Murine CENP-F Regulates Centrosomal Microtubule Nucleation and Interacts with Hook2 at the Centrosome
| Autor: | Ryan D. Pooley, Paul M. Miller, David M. Bader, Irina Kaverina, Katherine L. Moynihan |
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| Rok vydání: | 2009 |
| Předmět: |
Chromosomal Proteins
Non-Histone Cellular functions Centrosome cycle macromolecular substances Biology Microtubules Cell Line Mice Broad spectrum Microtubule Two-Hybrid System Techniques Chlorocebus aethiops Animals Humans Molecular Biology Microtubule nucleation Centrosome Mice Knockout Nocodazole Microfilament Proteins Articles Cell Biology Tubulin Modulators Cell biology COS Cells Microtubule-Associated Proteins Microtubule-Organizing Center |
| Zdroj: | Molecular Biology of the Cell. 20:4790-4803 |
| ISSN: | 1939-4586 1059-1524 |
| Popis: | The microtubule (MT) network is essential in a broad spectrum of cellular functions. Many studies have linked CENP-F to MT-based activities as disruption of this protein leads to major changes in MT structure and function. Still, the basis of CENP-F regulation of the MT network remains elusive. Here, our studies reveal a novel and critical localization and role for CENP-F at the centrosome, the major MT organizing center (MTOC) of the cell. Using a yeast two-hybrid screen, we identify Hook2, a linker protein that is essential for regulation of the MT network at the centrosome, as a binding partner of CENP-F. With recently developed immunochemical reagents, we confirm this interaction and reveal the novel localization of CENP-F at the centrosome. Importantly, in this first report of CENP-F−/−cells, we demonstrate that ablation of CENP-F protein function eliminates MT repolymerization after standard nocodazole treatment. This inhibition of MT regrowth is centrosome specific because MT repolymerization is readily observed from the Golgi in CENP-F−/−cells. The centrosome-specific function of CENP-F in the regulation of MT growth is confirmed by expression of truncated CENP-F containing only the Hook2-binding domain. Furthermore, analysis of partially reconstituted MTOC asters in cells that escape complete repolymerization block shows that disruption of CENP-F function impacts MT nucleation and anchoring rather than promoting catastrophe. Our study reveals a major new localization and function of CENP-F at the centrosome that is likely to impact a broad array of MT-based actions in the cell. |
| Databáze: | OpenAIRE |
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