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Tea4-PP1标志性结构通过双重募集Cdc42鸟嘌呤核苷酸交换因子(GEF)和排除GTP酶激活蛋白(GAP)来促进局部生长。

The Tea4-PP1 landmark promotes local growth by dual Cdc42 GEF recruitment and GAP exclusion.

作者信息

Kokkoris Kyriakos, Gallo Castro Daniela, Martin Sophie G

机构信息

University of Lausanne, Department of Fundamental Microbiology, Biophore Building, CH-1015 Lausanne, Switzerland.

出版信息

J Cell Sci. 2014 May 1;127(Pt 9):2005-16. doi: 10.1242/jcs.142174. Epub 2014 Feb 19.

Abstract

Cell polarization relies on small GTPases, such as Cdc42, which can break symmetry through self-organizing principles, and landmarks that define the axis of polarity. In fission yeast, microtubules deliver the Tea1-Tea4 complex to mark cell poles for growth, but how this complex activates Cdc42 is unknown. Here, we show that ectopic targeting of Tea4 to cell sides promotes the local activation of Cdc42 and cell growth. This activity requires that Tea4 binds the type I phosphatase (PP1) catalytic subunit Dis2 or Sds21, and ectopic targeting of either catalytic subunit is similarly instructive for growth. The Cdc42 guanine-nucleotide-exchange factor Gef1 and the GTPase-activating protein Rga4 are required for Tea4-PP1-dependent ectopic growth. Gef1 is recruited to ectopic Tea4 and Dis2 locations to promote Cdc42 activation. By contrast, Rga4 is locally excluded by Tea4, and its forced colocalization with Tea4 blocks ectopic growth, indicating that Rga4 must be present, but at sites distinct from Tea4. Thus, a Tea4-PP1 landmark promotes local Cdc42 activation and growth both through Cdc42 GEF recruitment and by creating a local trough in a Cdc42 GAP.

摘要

细胞极化依赖于小GTP酶,如Cdc42,其可通过自组织原理打破对称性,以及定义极性轴的标志物。在裂殖酵母中,微管将Tea1 - Tea4复合物输送至细胞极以标记生长位点,但该复合物如何激活Cdc42尚不清楚。在此,我们表明将Tea4异位靶向细胞侧面可促进Cdc42的局部激活和细胞生长。该活性要求Tea4结合I型磷酸酶(PP1)催化亚基Dis2或Sds21,且任一催化亚基的异位靶向对生长同样具有指导作用。Tea4 - PP1依赖性异位生长需要Cdc42鸟嘌呤核苷酸交换因子Gef1和GTP酶激活蛋白Rga4。Gef1被招募至异位的Tea4和Dis2位点以促进Cdc42激活。相比之下,Rga4被Tea4局部排除,其与Tea4的强制共定位会阻断异位生长,这表明Rga4必须存在,但要位于与Tea4不同的位点。因此,Tea4 - PP1标志物通过招募Cdc42鸟嘌呤核苷酸交换因子以及在Cdc42 GAP中形成局部低谷来促进局部Cdc42激活和生长。

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