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Published online 3 October 2005. doi:10.1083/jcb.200504107
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 171, Number 1, 27-33
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Rootletin forms centriole-associated filaments and functions in centrosome cohesion



Susanne Bahe1, York-Dieter Stierhof2, Christopher J. Wilkinson1, Florian Leiss1, and Erich A. Nigg1

1 Department of Cell Biology, Max-Planck-Institute for Biochemistry, D-82152 Martinsried, Germany
2 Electron Microscopy Unit, Center for Plant Molecular Biology, University of Tübingen, D-72076 Tübingen, Germany

Correspondence to Erich A. Nigg: nigg{at}biochem.mpg.de


Abstract

After duplication of the centriole pair during S phase, the centrosome functions as a single microtubule-organizing center until the onset of mitosis, when the duplicated centrosomes separate for bipolar spindle formation. The mechanisms regulating centrosome cohesion and separation during the cell cycle are not well understood. In this study, we analyze the protein rootletin as a candidate centrosome linker component. As shown by immunoelectron microscopy, endogenous rootletin forms striking fibers emanating from the proximal ends of centrioles. Moreover, rootletin interacts with C-Nap1, a protein previously implicated in centrosome cohesion. Similar to C-Nap1, rootletin is phosphorylated by Nek2 kinase and is displaced from centrosomes at the onset of mitosis. Whereas the overexpression of rootletin results in the formation of extensive fibers, small interfering RNA–mediated depletion of either rootletin or C-Nap1 causes centrosome splitting, suggesting that both proteins contribute to maintaining centrosome cohesion. The ability of rootletin to form centriole-associated fibers suggests a dynamic model for centrosome cohesion based on entangling filaments rather than continuous polymeric linkers.

C.J. Wilkinson's present address is Dept. of Anatomy, University of Cambridge, CB2 3DY Cambridge, UK.

F. Leiss's present address is Dept. of Molecular Neurobiology, Max Planck Institute of Neurobiology, D-82152 Martinsried, Germany.

Abbreviations used in this paper: IF, immunofluorescence; MT, microtubule; PCM, pericentriolar material; siRNA, small interfering RNA.


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