PMID: 9171381Jun 1, 1997Paper

Delineation of the interaction domains of Agrobacterium tumefaciens VirB7 and VirB9 by use of the yeast two-hybrid assay

Journal of Bacteriology
Anath DasY H Xie

Abstract

The Agrobacterium tumefaciens VirB proteins are postulated to form a transport pore for the transfer of T-DNA. Formation of the transport pore will involve interactions among the VirB proteins. A powerful genetic method to study protein-protein interaction is the yeast two-hybrid assay. To test whether this method can be used to study interactions among the VirB membrane proteins, we studied the interaction of VirB7 and VirB9 in yeast. We recently demonstrated that VirB7 and VirB9 form a protein complex linked by a disulfide bond between cysteine 24 of VirB7 and cysteine 262 of VirB9 (L. Anderson, A. Hertzel, and A. Das, Proc. Natl. Acad. Sci. USA 93:8889-8894, 1996). We now demonstrate that VirB7 and VirB9 interact in yeast, and this interaction does not require the cysteine residues essential for the disulfide linkage. By using defined segments in fusion constructions, we mapped the VirB7 interaction domain of VirB9 to residues 173 to 275. In tumor formation assays, both virB7C24S and virB9C262S expressed from a multicopy plasmid complemented the respective deletion mutation, indicating that the cysteine residues may not be essential for DNA transfer.

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Citations

Jun 17, 1998·Proceedings of the National Academy of Sciences of the United States of America·W DengE W Nester
Sep 13, 2005·Annual Review of Microbiology·Peter J ChristieEric Cascales
Dec 5, 2008·Molecular Microbiology·Simon J JakubowskiPeter J Christie
Aug 2, 2020·Molecular Microbiology·Birgit KochNatalio Krasnogor
Mar 24, 2004·Nature Reviews. Microbiology·Eric Cascales, Peter J Christie
Sep 17, 2009·Nature Reviews. Microbiology·Rémi FronzesGabriel Waksman
Aug 15, 2002·Proceedings of the National Academy of Sciences of the United States of America·Doyle V WardPatricia C Zambryski
Aug 6, 2000·The Plant Journal : for Cell and Molecular Biology·J ZupanP Zambryski
Aug 20, 2003·Proceedings of the National Academy of Sciences of the United States of America·Matxalen LlosaFernando de la Cruz
Jan 30, 2013·Biochemical Society Transactions·Vidya Chandran
Sep 22, 2001·Journal of Bacteriology·E SagulenkoP J Christie
Jul 3, 2004·Journal of Bacteriology·Alireza Shamaei-TousiGad Frankel
May 24, 2001·Journal of Bacteriology·V SagulenkoP J Christie
Jun 22, 2006·Journal of Bacteriology·Valerie J BuslerTimothy L Cover
Jul 12, 2003·FEMS Microbiology Letters·T D LawleyL S Frost
Dec 3, 2014·Biochemical and Biophysical Research Communications·Gopal Jee GopalGauranga Mukhopadhyay
Jul 19, 2003·International Journal of Medical Microbiology : IJMM·Dirk HofreuterRainer Haas

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