PMID: 8943032Nov 26, 1996Paper

A positive genetic selection for disrupting protein-protein interactions: identification of CREB mutations that prevent association with the coactivator CBP

Proceedings of the National Academy of Sciences of the United States of America
H M ShihM F Hoekstra

Abstract

The Escherichia coli tet-repressor (TetR) operator system was used to develop a variation of the yeast two-hybrid assay in which disruptions of protein-protein interactions can be identified by a positive selection. This assay, designated the "split-hybrid system," contains a two-component reporter. The first component contains LexA binding sites upstream of the TetR gene and the second contains TetR operator binding sites upstream of HIS3. Interaction of one protein fused to the LexA DNA binding domain with a second protein fused to the VP16 activation domain results in TetR expression. TetR subsequently binds to the tet operators, blocking the expression of HIS3 and preventing yeast growth in media lacking histidine. The utility of the split-hybrid system was analyzed by examining the phosphorylation-dependent interaction of CREB and its coactivator CREB binding protein (CBP). CREB and CBP associate through an interaction that depends upon CREB phosphorylation at Ser-133. Mutation of this phosphorylation site prevents yeast growth in the standard two-hybrid assay but allows growth in the split-hybrid strains. The split-hybrid system was used to identify other CREB mutations that disrupt its association with CBP. These mutatio...Continue Reading

References

Jan 1, 1991·Methods in Enzymology·R S Sikorski, J D Boeke
Sep 1, 1990·Trends in Biochemical Sciences·B E Kemp, R B Pearson
Nov 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·M FujiiI M Verma
Jan 1, 1994·Annual Review of Microbiology·W Hillen, C Berens
Oct 28, 1993·Nature·J C ChriviaR H Goodman
Apr 30, 1996·Proceedings of the National Academy of Sciences of the United States of America·R K BrachmannJ D Boeke

❮ Previous
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Citations

Aug 28, 1998·Trends in Biotechnology·P Colas, R Brent
Mar 21, 1998·Current Opinion in Biotechnology·R M Frederickson
Jun 11, 2002·Methods : a Companion to Methods in Enzymology·John D Crispino, Stuart H Orkin
Feb 13, 2001·Current Opinion in Chemical Biology·S J BoultonM Vidal
Feb 18, 1999·Current Opinion in Chemical Biology·B L Drees
Oct 27, 2009·Nature Methods·Matija DrezeMarc Vidal
Nov 8, 2013·Nature Reviews. Genetics·Colm J RyanNevan J Krogan
Nov 18, 2000·AIDS Research and Human Retroviruses·Y L KuoC Z Giam
Aug 29, 2002·The EMBO Journal·Fran LedoJose R Naranjo
Feb 12, 2000·Molecular and Cellular Biology·J R CardinauxR H Goodman
Mar 22, 2001·Molecular and Cellular Biology·P ErnstS J Korsmeyer
Jun 29, 2000·Annual Review of Biochemistry·A J Shaywitz, M E Greenberg
Jan 1, 1997·Annual Review of Genetics·R Brent, R L Finley
May 8, 2003·Biological Procedures Online·Wim Van Criekinge, Rudi Beyaert
Nov 15, 2003·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Michael FetchkoIgor Stagljar
Feb 12, 1998·Proceedings of the National Academy of Sciences of the United States of America·J Huang, S L Schreiber
Jul 19, 2002·Reviews in Medical Virology·Arianna LoregianGiorgio Palù
Jul 1, 2009·The Plant Journal : for Cell and Molecular Biology·Ayalew LigabaMiguel Piñeros
Feb 6, 2004·Journal of Biochemistry and Molecular Biology·Sayeon ChoByoung Chul Park
Mar 20, 2008·Expert Opinion on Therapeutic Targets·Nicolas Lentze, Daniel Auerbach
May 22, 2001·Medical Hypotheses·B SainzJ M Hill
Jul 3, 2003·Chemistry & Biology·Jason W ChinPeter G Schultz
Jun 10, 2014·Journal of Chemical Theory and Computation·Mateusz KurcinskiSebastian Kmiecik
Dec 8, 2006·Molecular & Cellular Proteomics : MCP·Phillip N GrayThomas G Chappell
Oct 19, 2017·Nature Methods·Jonathan WoodsmithUlrich Stelzl
Dec 21, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·C T YuM Z Lai
Jan 5, 2002·The Journal of Biological Chemistry·Joanna R MorrisEllen Solomon

❮ Previous
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