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A fluorescent two-hybrid assay for direct visualization of protein interactions in living cells.

Zolghadr, Kourosh ; Mortusewicz, Oliver ; Rothbauer, Ulrich ; Kleinhans, Regina ; Goehler, Heike ; Wanker, Erich E. ; Cardoso, M. Cristina ; Leonhardt, Heinrich (2008):
A fluorescent two-hybrid assay for direct visualization of protein interactions in living cells.
In: Molecular & cellular proteomics : MCP, 7 (11), pp. 2279-87. ISSN 1535-9484,
[Article]

Abstract

Genetic high throughput screens have yielded large sets of potential protein-protein interactions now to be verified and further investigated. Here we present a simple assay to directly visualize protein-protein interactions in single living cells. Using a modified lac repressor system, we tethered a fluorescent bait at a chromosomal lac operator array and assayed for co-localization of fluorescent prey fusion proteins. With this fluorescent two-hybrid assay we successfully investigated the interaction of proteins from different subcellular compartments including nucleus, cytoplasm, and mitochondria. In combination with an S phase marker we also studied the cell cycle dependence of protein-protein interactions. These results indicate that the fluorescent two-hybrid assay is a powerful tool to investigate protein-protein interactions within their cellular environment and to monitor the response to external stimuli in real time.

Item Type: Article
Erschienen: 2008
Creators: Zolghadr, Kourosh ; Mortusewicz, Oliver ; Rothbauer, Ulrich ; Kleinhans, Regina ; Goehler, Heike ; Wanker, Erich E. ; Cardoso, M. Cristina ; Leonhardt, Heinrich
Title: A fluorescent two-hybrid assay for direct visualization of protein interactions in living cells.
Language: English
Abstract:

Genetic high throughput screens have yielded large sets of potential protein-protein interactions now to be verified and further investigated. Here we present a simple assay to directly visualize protein-protein interactions in single living cells. Using a modified lac repressor system, we tethered a fluorescent bait at a chromosomal lac operator array and assayed for co-localization of fluorescent prey fusion proteins. With this fluorescent two-hybrid assay we successfully investigated the interaction of proteins from different subcellular compartments including nucleus, cytoplasm, and mitochondria. In combination with an S phase marker we also studied the cell cycle dependence of protein-protein interactions. These results indicate that the fluorescent two-hybrid assay is a powerful tool to investigate protein-protein interactions within their cellular environment and to monitor the response to external stimuli in real time.

Journal or Publication Title: Molecular & cellular proteomics : MCP
Journal volume: 7
Number: 11
Divisions: 10 Department of Biology > Cell Biology and Epigenetics
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10 Department of Biology
Date Deposited: 06 Mar 2010 16:06
Official URL: http://www.cardoso-lab.org/publications/Zolghadr_2008.pdf
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