Probing Cdc42 Polarization Dynamics in Budding Yeast Using a Biosensor

Satoshi Okada, Mid Eum Lee, Erfei Bi, Hay Oak Park

研究成果: Chapter in Book/Report/Conference proceedingChapter

8 被引用数 (Scopus)

抄録

Cdc42 is a small guanosine triphosphatase (GTPase) that plays a central role in polarity development in diverse cell types. Since the activity of Cdc42 is dynamically controlled in time and space, it is required to develop a biosensor to monitor its activation in vivo. In this chapter, we describe the construction and usage of a simple and robust biosensor for monitoring active Cdc42 in budding yeast. This affinity-based biosensor uses a red fluorescent protein fused to a Cdc42- and Rac-interactive binding motif from one of the Cdc42 effector proteins. Because it binds specifically to the GTP-bound Cdc42, this biosensor can be used to monitor Cdc42 activation in vivo. This or similar biosensors can be widely used for studying GTPase signaling in other cell types because of the conserved CRIB motif present among GTPase targets.

本文言語英語
ホスト出版物のタイトルMethods in Enzymology
出版社Academic Press Inc.
ページ171-190
ページ数20
DOI
出版ステータス出版済み - 2017
外部発表はい

出版物シリーズ

名前Methods in Enzymology
589
ISSN(印刷版)0076-6879
ISSN(電子版)1557-7988

All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学

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