Genetically encoded photoswitching of actin assembly through the Cdc42-WASP-Arp2/3 complex pathway

Daisy W. Leung, Chinatsu Otomo, Joanne Chory, Michael K. Rosen

Research output: Contribution to journalArticlepeer-review

78 Scopus citations

Abstract

General methods to engineer genetically encoded, reversible, light-mediated control over protein function would be useful in many areas of biomedical research and technology. We describe a system that yields such photo-control over actin assembly. We fused the Rho family GTPase Cdc42 in its GDP-bound form to the photosensory domain of phytochrome B (PhyB) and fused the Cdc42 effector, the Wiskott-Aldrich Syndrome Protein (WASP), to the light-dependent PhyB-binding domain of phytochrome interacting factor 3 (Pif3). Upon red light illumination, the fusion proteins bind each other, activating WASP, and consequently stimulating actin assembly by the WASP target, the Arp2/3 complex. Binding and WASP activation are reversed by far-red illumination. Our approach, in which the biochemical specificity of the nucleotide switch in Cdc42 is overridden by the light-dependent PhyB-Pif3 interaction, should be generally applicable to other GTPase-effector pairs.

Original languageEnglish (US)
Pages (from-to)12797-12802
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume105
Issue number35
DOIs
StatePublished - Sep 2 2008

Keywords

  • Cytoskeleton
  • Photoswitch
  • Phytochrome
  • Protein engineering
  • Signal transduction

ASJC Scopus subject areas

  • General

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