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Published online 27 December 2000. doi:10.1083/jcb.151.7.1525
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© The Rockefeller University Press, 0021-9525/2000/12/1525/ $5.00
The Journal of Cell Biology, Volume 151, Number 7, December 25, 2000 1525-1536

Original Article

The Role of Xgrip210 in {gamma}-Tubulin Ring Complex Assembly and Centrosome Recruitment

Lijun Zhanga, Thomas J. Keatingb, Andrew Wildea, Gary G. Borisyb, and Yixian Zhenga
a Howard Hughes Medical Institute, Carnegie Institution of Washington, Baltimore, Maryland 21210
b Laboratory of Molecular Biology, University of Wisconsin, Madison, Wisconsin 53706

Correspondence to: Yixian Zheng, Howard Hughes Medical Institute, Carnegie Institution of Washington, 115 West University Parkway, Baltimore, MD 21210. Tel:(410) 554-1232 Fax:(410) 243-6311 E-mail:zheng{at}ciwemb.edu.

The {gamma}-tubulin ring complex ({gamma}TuRC), purified from the cytoplasm of vertebrate and invertebrate cells, is a microtubule nucleator in vitro. Structural studies have shown that {gamma}TuRC is a structure shaped like a lock-washer and topped with a cap. Microtubules are thought to nucleate from the uncapped side of the {gamma}TuRC. Consequently, the cap structure of the {gamma}TuRC is distal to the base of the microtubules, giving the end of the microtubule the shape of a pointed cap. Here, we report the cloning and characterization of a new subunit of Xenopus {gamma}TuRC, Xgrip210. We show that Xgrip210 is a conserved centrosomal protein that is essential for the formation of {gamma}TuRC. Using immunogold labeling, we found that Xgrip210 is localized to the ends of microtubules nucleated by the {gamma}TuRC and that its localization is more distal, toward the tip of the {gamma}TuRC-cap structure, than that of {gamma}-tubulin. Immunodepletion of Xgrip210 blocks not only the assembly of the {gamma}TuRC, but also the recruitment of {gamma}-tubulin and its interacting protein, Xgrip109, to the centrosome. These results suggest that Xgrip210 is a component of the {gamma}TuRC cap structure that is required for the assembly of the {gamma}TuRC.

Key Words: Xgrip210, centrosome, microtubule nucleation, {gamma}-tubulin ring complex, Xenopus


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