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A Genetic System Based on Split-Ubiquitin for the Analysis of Interactions between Membrane Proteins in vivo
A detection system for interactions between membrane proteins in vivo is described. The system is based on split-ubiquitin [Johnson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein...
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Published in: | Proceedings of the National Academy of Sciences - PNAS 1998-04, Vol.95 (9), p.5187-5192 |
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creator | Stagljar, Igor Korostensky, Chantal Johnsson, Nils Heesen, Stephan Te |
description | A detection system for interactions between membrane proteins in vivo is described. The system is based on split-ubiquitin [Johnson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein fusion. The cleavage releases a transcription factor, which activates reporter genes in the nucleus. As a result, interaction between membrane proteins can be analyzed by the means of a colorimetric assay. We use membrane proteins of the endoplasmic reticulum as a model system. Wbp1p and Ost1p are both subunits of the oligosaccharyl transferase membrane protein complex. The Alg5 protein also localizes to the membrane of the endoplasmic reticulum, but does not interact with the oligosaccharyltransferase. Specific interactions are detected between Wbp1p and Ost1p, but not between Wbp1p and Alg5p. The new system might be useful as a genetic and biochemical tool for the analysis of interactions between membrane proteins in vivo. |
doi_str_mv | 10.1073/pnas.95.9.5187 |
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The system is based on split-ubiquitin [Johnson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein fusion. The cleavage releases a transcription factor, which activates reporter genes in the nucleus. As a result, interaction between membrane proteins can be analyzed by the means of a colorimetric assay. We use membrane proteins of the endoplasmic reticulum as a model system. Wbp1p and Ost1p are both subunits of the oligosaccharyl transferase membrane protein complex. The Alg5 protein also localizes to the membrane of the endoplasmic reticulum, but does not interact with the oligosaccharyltransferase. Specific interactions are detected between Wbp1p and Ost1p, but not between Wbp1p and Alg5p. The new system might be useful as a genetic and biochemical tool for the analysis of interactions between membrane proteins in vivo.</description><subject>Amino acids</subject><subject>Biological Sciences</subject><subject>Cell growth</subject><subject>Cloning, Molecular - methods</subject><subject>Endopeptidases - metabolism</subject><subject>Genetic Vectors</subject><subject>Genetics</subject><subject>Herpes Simplex Virus Protein Vmw65 - metabolism</subject><subject>Hexosyltransferases</subject><subject>Journalism</subject><subject>Membrane proteins</subject><subject>Membrane Proteins - chemistry</subject><subject>Membranes</subject><subject>Plasmids</subject><subject>Protein Binding</subject><subject>Proteins</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Reporter genes</subject><subject>Saccharomyces cerevisiae</subject><subject>Transferases - metabolism</subject><subject>Ubiquitin-Specific Proteases</subject><subject>Ubiquitins</subject><subject>Ubiquitins - chemistry</subject><subject>Yeasts</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><recordid>eNqFkcFv0zAUhyMEGmVw5YCEZHHYLeU5sZ1Y4tJNMCYNgTR2tpz0mblK7c52Cv3vcdSqGxzg5MPv-579_CuK1xTmFJr6_cbpOJd8Luects2TYkZB0lIwCU-LGUDVlC2r2PPiRYwrAJC8hZPiRHIBFaezYliQS3SYbE9udjHhmpzriEviHbnZDDaVt529H22yjhgfSLpDsnB62EUbiTfkyiUMuk_Wu0g6TD8RHfmC6y5oh-Rb8AltTrK9tVv_snhm9BDx1eE8LW4_ffx-8bm8_np5dbG4LntO61TKrqdMs77moq2XEoyh-eEMTQcGG-SGAyBruEa5bETX99o0tTBARQtLZKw-LT7s527Gbo3LHl0KelCbYNc67JTXVv2ZOHunfvitqqCqRdbPDnrw9yPGpNY29jgMeSc_RtXItqqE-D9IRc2Bsgl89xe48mPI_xjzlbQWeSJkaL6H-uBjDGiOD6agpq7V1LWSXEk1dZ2Ft4_XPOKHch_lk_eQPvhn_8qVGYch4a-UwTd7cBWTD0eSMdHK-jf7EseB</recordid><startdate>19980428</startdate><enddate>19980428</enddate><creator>Stagljar, Igor</creator><creator>Korostensky, Chantal</creator><creator>Johnsson, Nils</creator><creator>Heesen, Stephan Te</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19980428</creationdate><title>A Genetic System Based on Split-Ubiquitin for the Analysis of Interactions between Membrane Proteins in vivo</title><author>Stagljar, Igor ; Korostensky, Chantal ; Johnsson, Nils ; Heesen, Stephan Te</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c513t-9bc14a4c35683d90ff10094efb0fe7e5f500e475ae9d76bccaf736f01680de443</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Amino acids</topic><topic>Biological Sciences</topic><topic>Cell growth</topic><topic>Cloning, Molecular - methods</topic><topic>Endopeptidases - metabolism</topic><topic>Genetic Vectors</topic><topic>Genetics</topic><topic>Herpes Simplex Virus Protein Vmw65 - metabolism</topic><topic>Hexosyltransferases</topic><topic>Journalism</topic><topic>Membrane proteins</topic><topic>Membrane Proteins - chemistry</topic><topic>Membranes</topic><topic>Plasmids</topic><topic>Protein Binding</topic><topic>Proteins</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Reporter genes</topic><topic>Saccharomyces cerevisiae</topic><topic>Transferases - metabolism</topic><topic>Ubiquitin-Specific Proteases</topic><topic>Ubiquitins</topic><topic>Ubiquitins - chemistry</topic><topic>Yeasts</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Stagljar, Igor</creatorcontrib><creatorcontrib>Korostensky, Chantal</creatorcontrib><creatorcontrib>Johnsson, Nils</creatorcontrib><creatorcontrib>Heesen, Stephan Te</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Stagljar, Igor</au><au>Korostensky, Chantal</au><au>Johnsson, Nils</au><au>Heesen, Stephan Te</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Genetic System Based on Split-Ubiquitin for the Analysis of Interactions between Membrane Proteins in vivo</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1998-04-28</date><risdate>1998</risdate><volume>95</volume><issue>9</issue><spage>5187</spage><epage>5192</epage><pages>5187-5192</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><notes>ObjectType-Article-2</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-1</notes><notes>content type line 23</notes><notes>ObjectType-Article-1</notes><notes>ObjectType-Feature-2</notes><notes>Present address: Department Informatik, Haldeneggsteig 4, Eidgenössiche Technische Hochschule Zürich, CH-8092 Zürich, Switzerland.</notes><notes>To whom reprint requests should be addressed. e-mail: teheesen@micro.biol.ethz.ch.</notes><notes>Edited by Alexander Varshavsky, California Institute of Technology, Pasadena, CA, and approved February 22, 1998</notes><abstract>A detection system for interactions between membrane proteins in vivo is described. The system is based on split-ubiquitin [Johnson, N. & Varshavsky, A. (1994) Proc. Natl. Acad. Sci. USA 91, 10340-10344]. Interaction between two membrane proteins is detected by proteolytic cleavage of a protein fusion. The cleavage releases a transcription factor, which activates reporter genes in the nucleus. As a result, interaction between membrane proteins can be analyzed by the means of a colorimetric assay. We use membrane proteins of the endoplasmic reticulum as a model system. Wbp1p and Ost1p are both subunits of the oligosaccharyl transferase membrane protein complex. The Alg5 protein also localizes to the membrane of the endoplasmic reticulum, but does not interact with the oligosaccharyltransferase. Specific interactions are detected between Wbp1p and Ost1p, but not between Wbp1p and Alg5p. The new system might be useful as a genetic and biochemical tool for the analysis of interactions between membrane proteins in vivo.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>9560251</pmid><doi>10.1073/pnas.95.9.5187</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Amino acids Biological Sciences Cell growth Cloning, Molecular - methods Endopeptidases - metabolism Genetic Vectors Genetics Herpes Simplex Virus Protein Vmw65 - metabolism Hexosyltransferases Journalism Membrane proteins Membrane Proteins - chemistry Membranes Plasmids Protein Binding Proteins Recombinant Fusion Proteins - metabolism Reporter genes Saccharomyces cerevisiae Transferases - metabolism Ubiquitin-Specific Proteases Ubiquitins Ubiquitins - chemistry Yeasts |
title | A Genetic System Based on Split-Ubiquitin for the Analysis of Interactions between Membrane Proteins in vivo |
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