Witte Kristen, Strickland Devin, Glotzer Michael
Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, United States.
Elife. 2017 Jul 6;6:e26722. doi: 10.7554/eLife.26722.
Cell polarization underlies many cellular and organismal functions. The GTPase Cdc42 orchestrates polarization in many contexts. In budding yeast, polarization is associated with a focus of Cdc42•GTP which is thought to self sustain by recruiting a complex containing Cla4, a Cdc42-binding effector, Bem1, a scaffold, and Cdc24, a Cdc42 GEF. Using optogenetics, we probe yeast polarization and find that local recruitment of Cdc24 or Bem1 is sufficient to induce polarization by triggering self-sustaining Cdc42 activity. However, the response to these perturbations depends on the recruited molecule, the cell cycle stage, and existing polarization sites. Before cell cycle entry, recruitment of Cdc24, but not Bem1, induces a metastable pool of Cdc42 that is sustained by positive feedback. Upon Cdk1 activation, recruitment of either Cdc24 or Bem1 creates a stable site of polarization that induces budding and inhibits formation of competing sites. Local perturbations have therefore revealed unexpected features of polarity establishment.
细胞极化是许多细胞和机体功能的基础。GTP酶Cdc42在多种情况下协调极化过程。在芽殖酵母中,极化与Cdc42•GTP的一个焦点相关,该焦点被认为通过招募一个包含Cla4(一种Cdc42结合效应器)、Bem1(一种支架蛋白)和Cdc24(一种Cdc42鸟苷酸交换因子)的复合物来实现自我维持。利用光遗传学技术,我们对酵母极化进行了探究,发现局部招募Cdc24或Bem1足以通过触发自我维持的Cdc42活性来诱导极化。然而,对这些扰动的反应取决于所招募的分子、细胞周期阶段以及现有的极化位点。在细胞周期进入之前,招募Cdc24而非Bem1会诱导一个由正反馈维持的Cdc42亚稳态池。在Cdk1激活后,招募Cdc24或Bem1都会产生一个稳定的极化位点,该位点诱导出芽并抑制竞争位点的形成。因此,局部扰动揭示了极性建立的意外特征。