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Journal of Bacteriology, February 2001, p. 1022-1031, Vol. 183, No. 3
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.3.1022-1031.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Cooperativity between KorB and TrbA Repressors of Broad-Host-Range Plasmid RK2

Malgorzata Zatyka,dagger Lewis Bingle, Anthony C. Jones, and Christopher M. Thomas*

School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom

Received 9 May 2000/Accepted 5 November 2000

The KorB and TrbA proteins of broad-host-range plasmid RK2 are key regulators of the plasmid genes required for conjugative transfer. trbBp is the primary promoter responsible for expression of mating pair formation genes. We show that despite the targets for KorB and TrbA at trbBp being about 165 bp apart, 189 bp upstream of the transcription start point and overlapping the -10 region, respectively, these two proteins show up to 10-fold cooperativity for the repression of trbBp. Deletion analysis of TrbA showed that the C-terminal domain (CTD), which has a high degree of sequence conservation with the CTD of KorA, is required for this cooperativity with KorB. Western blotting demonstrated that the apparently mutual enhancement of repression is not due simply to elevation of repressor level by the presence of the second protein, suggesting that the basis for cooperativity is interaction between KorB and TrbA bound at their respective operators.

* Corresponding author. Mailing address: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom. Phone: 0121 414 5903. Fax: 0121 414 5925. E-mail: c.m.thomas{at}bham.ac.uk.

dagger Present address: Department of Paediatrics & Child Health, Division of Medical & Molecular Genetics, The Medical School, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.

Journal of Bacteriology, February 2001, p. 1022-1031, Vol. 183, No. 3
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.3.1022-1031.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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