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A more recent version of this article appeared on January 1, 2008
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Submitted on September 10, 2007
Revised on October 12, 2007
Accepted on October 18, 2007
-Tubulin Small Complex: Implications of Its Architecture and Flexibility for Microtubule Nucleation
*Department of Biochemistry and Biophysics and the Howard Hughes Medical Institute, University of California, San Francisco, CA 94158;
Department of Biochemistry, University of Washington, Seattle, WA 98195
Monitoring Editor: Tim Stearns
The
-tubulin small complex (
-TuSC) is an evolutionarily conserved heterotetramer essential for microtubule nucleation. We have determined the structure of the S. cerevisiae
-TuSC at 25 Å resolution by electron microscopy.
-TuSC is Y-shaped, with an elongated body connected to two arms. Gold labeling showed that the two
-tubulins are located in lobes at the ends of the arms, and the relative orientations of the other
-TuSC components were determined by in vivo FRET. The structures of different subpopulations of
-TuSC indicate flexibility in the connection between a mobile arm and the rest of the complex, resulting in variation of the relative positions and orientations of the
-tubulins. In all of the structures the
-tubulins are distinctly separated, a configuration incompatible with the microtubule lattice. The separation of the
-tubulins in isolated
-TuSC likely plays a role in suppressing its intrinsic microtubule nucleating activity, which is relatively weak until the
-TuSC is incorporated into higher-order complexes or localized to microtubule organizing centers. We propose that further movement of the mobile arm is required to bring the
-tubulins together in microtubule-like interactions, and provide a template for microtubule growth.