Bioluminescence resonance energy transfer identify scaffold protein CNK1 interactions in intact cells
| Autor: | Soon-Young Na, Julio C. Bernabe-Ortiz, Marco Lopez-Ilasaca, Ramnik J. Xavier, Victor J. Dzau |
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| Rok vydání: | 2004 |
| Předmět: |
Scaffold protein
RHOA Biophysics Bioluminescence resonance energy transfer Biochemistry Cell Line Mice Rho Structural Biology In vivo Genetics Animals Humans Immunoprecipitation Enhancer Molecular Biology Cells Cultured Adaptor Proteins Signal Transducing biology Cell Biology Actin cytoskeleton In vitro Cell biology Pleckstrin homology domain Serum response element Energy Transfer Microscopy Fluorescence Luminescent Measurements hCNK1 biology.protein Signal transduction Ras Signal Transduction |
| Zdroj: | FEBS Letters. 579:648-654 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/j.febslet.2004.12.039 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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