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 |
| Volume of the journal: | 7 |
| Issue Number: | 11 |
| Divisions: | 10 Department of Biology > Cell Biology and Epigenetics ?? fb10_zoologie ?? 10 Department of Biology |
| Date Deposited: | 06 Mar 2010 16:06 |
| URL / URN: | http://www.cardoso-lab.org/publications/Zolghadr_2008.pdf |
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