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Coordination of Rho GTPase activities during cell protrusion

机译：细胞突起过程中Rho GTPase活性的协调

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摘要

The GTPases Racl, RhoA and Cdc42 act together to control cytos-keleton dynamics. Recent biosensor studies have shown that all three GTPases are activated at the front of migrating cells, and biochemical evidence suggests that they may regulate one another: Cdc42 can activate Racl (ref. 8), and Racl and RhoA are mutually inhibitory. However, their spatiotemporal coordination, at the seconds and single-micrometre dimensions typical of individual protrusion events, remains unknown. Here we examine GTPase coordination in mouse embryonic fibroblasts both through simultaneous visualization of two GTPase biosensors and using a 'computational multiplexing' approach capable of defining the relationships between multiple protein activities visualized in separate experiments. We found that RhoA is activated at the cell edge synchronous with edge advancement, whereas Cdc42 and Racl are activated 2 μm behind the edge with a delay of 40 s. This indicates that Racl and RhoA operate antagonistically through spatial separation and precise timing, and that RhoA has a role in the initial events of protrusion, whereas Racl and Cdc42 activate pathways implicated in reinforcement and stabilization of newly expanded protrusions.
机译：GTPases Racl，RhoA和Cdc42共同作用以控制细胞骨架的动力学。最近的生物传感器研究表明，所有三个GTPases都在迁移细胞的前面被激活，生化证据表明它们可能相互调节：Cdc42可以激活Racl（参考文献8），而Racl和RhoA具有相互抑制作用。然而，它们在时空上的协调性，在几秒钟内以及单个突出事件所特有的单微米尺寸，仍然未知。在这里，我们通过同时可视化两个GTPase生物传感器并使用能够定义在单独实验中可视化的多种蛋白质活性之间的关系的“计算多重化”方法，来检查小鼠胚胎成纤维细胞中的GTPase配位。我们发现，RhoA在细胞边缘与边缘推进同步激活，而Cdc42和Racl在边缘后2μm处激活，延迟40 s。这表明Rac1和RhoA通过空间分离和精确的时间拮抗作用，并且RhoA在突出的初始事件中起作用，而Rac1和Cdc42激活与新扩展的突出物的增强和稳定有关的途径。

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  • 来源
    《Nature》 |2009年第7260期|99-103|共5页
  • 作者

    Matthias Machacek; Louis Hodgson; Christopher Welch; Hunter Elliott; Olivier Pertz; Perihan Nalbant; Amy Abell; Gary L. Johnson; Klaus M. Hahn; Gaudenz Danuser;

  • 作者单位

    Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA Novartis Pharma AG, Lichtstrasse 35, CH-4056 Basel, Switzerland;

    Departments of Pharmacology, Medicinal Chemistry and Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA Department of Anatomy and Structural Biology and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Ave, Bronx, New York 10461, USA;

    Departments of Pharmacology, Medicinal Chemistry and Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA;

    Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA;

    Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA Department of Biomedicine, University of Basel, Mattenstrasse 28, CH-4058 Basel, Switzerland;

    Department of Molecular Cell Biology, Center for Medical Biotechnology, University of Duisburg-Essen, 45117 Essen, Germany;

    Departments of Pharmacology, Medicinal Chemistry and Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA;

    Departments of Pharmacology, Medicinal Chemistry and Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA;

    Departments of Pharmacology, Medicinal Chemistry and Lineberger Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA;

    Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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