Bioluminescence resonance energy transfer identify scaffold protein CNK1 interactions in intact cells.

Journal Article

Connector enhancer of KSR (CNK) proteins have been proposed to act as scaffolds in the Ras-MAPK pathway. In this work, using in vivo bioluminescence resonance energy transfer (BRET) assays and in vitro co-immunoprecipitation, we show that hCNK1 interacts with the active form of Rho A (G14V) proteins. The domain of hCNK1 that allows binding to Rho proteins involves the C-terminal PH domain. Overexpression of hCNK1 does not affect the actin cytoskeleton and does not modify the appearance of stress fibers in cells overexpressing a constitutively active form of RhoA. In contrast, hCNK1 was able to significantly decrease the RhoA-induced transcriptional activity of the serum response element (SRE) without effect on the Ras-induced SRE activation. These results identify hCNK1 as a specific partner of Rho proteins both in vitro and in vivo and suggest a role of hCNK1 in the signal transduction of Rho proteins.

Full Text

Duke Authors

Cited Authors

  • Lopez-Ilasaca, MA; Bernabe-Ortiz, JC; Na, S-Y; Dzau, VJ; Xavier, RJ

Published Date

  • January 31, 2005

Published In

Volume / Issue

  • 579 / 3

Start / End Page

  • 648 - 654

PubMed ID

  • 15670823

International Standard Serial Number (ISSN)

  • 0014-5793

Digital Object Identifier (DOI)

  • 10.1016/j.febslet.2004.12.039

Language

  • eng

Conference Location

  • England