Department of Biophysics, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390, USA.
Science. 2012 Aug 17;337(6096):857-60. doi: 10.1126/science.1221698.
Stu2p/XMAP215/Dis1 family proteins are evolutionarily conserved regulatory factors that use αβ-tubulin-interacting tumor overexpressed gene (TOG) domains to catalyze fast microtubule growth. Catalysis requires that these polymerases discriminate between unpolymerized and polymerized forms of αβ-tubulin, but the mechanism by which they do so has remained unclear. Here, we report the structure of the TOG1 domain from Stu2p bound to yeast αβ-tubulin. TOG1 binds αβ-tubulin in a way that excludes equivalent binding of a second TOG domain. Furthermore, TOG1 preferentially binds a curved conformation of αβ-tubulin that cannot be incorporated into microtubules, contacting α- and β-tubulin surfaces that do not participate in microtubule assembly. Conformation-selective interactions with αβ-tubulin explain how TOG-containing polymerases discriminate between unpolymerized and polymerized forms of αβ-tubulin and how they selectively recognize the growing end of the microtubule.
Stu2p/XMAP215/Dis1 家族蛋白是进化上保守的调节因子,它们使用αβ-微管相互作用肿瘤过表达基因(TOG)结构域来催化快速微管生长。催化作用需要这些聚合酶区分未聚合和聚合形式的αβ-微管,但它们的作用机制仍不清楚。在这里,我们报告了与酵母αβ-微管结合的 Stu2p 的 TOG1 结构域的结构。TOG1 以排除第二个 TOG 结构域等效结合的方式与αβ-微管结合。此外,TOG1 优先结合不能掺入微管的αβ-微管的弯曲构象,与不参与微管组装的α-和β-微管表面接触。与αβ-微管的构象选择性相互作用解释了含有 TOG 的聚合酶如何区分αβ-微管的未聚合和聚合形式,以及它们如何选择性地识别微管的生长端。