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Proc Natl Acad Sci U S A. 2008 Sep 2;105(35):12797-802. doi: 10.1073/pnas.0801232105. Epub 2008 Aug 26.

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

Author information

1
Howard Hughes Medical Institute, Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.

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.

PMID:
18728185
PMCID:
PMC2525560
DOI:
10.1073/pnas.0801232105
[Indexed for MEDLINE]
Free PMC Article

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