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Journal of Bacteriology, March 2003, p. 1817-1824, Vol. 185, No. 6
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.6.1817-1824.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of a Novel Membrane-Associated Gene Product That Suppresses Toxicity of a TrfA Peptide from Plasmid RK2 and Its Relationship to the DnaA Host Initiation Protein

Peter D. Kim,1 Trevor Banack,1 Daniel M. Lerman,1 Jeremiah C. Tracy,1 Johanna Eltz Camara,2 Elliot Crooke,2 Don Oliver,1 and William Firshein1*

Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, Connecticut 06459,1 Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, Washington, D.C. 200072

Received 11 October 2002/ Accepted 30 December 2002

The toxicity of a peptide derived from the amino-terminal portion of 33-kDa TrfA, one of the initiation proteins encoded by the broad-host-range plasmid RK2, was suppressed by a host protein related to DnaA, the initiation protein of Escherichia coli. The newly identified 28.4-kDa protein, termed a DnaA paralog (Dp) because it is similar to a region of DnaA but likely has a different function in initiation of plasmid RK2 replication, interacts physically with the 33-kDa TrfA initiation protein, including the initiation-active monomeric form. The Dp has a cellular distribution similar to that of the 33-kDa TrfA initiation protein, being found primarily in the inner membrane fraction, with lesser amounts detected in the outer membrane fraction and almost none in the soluble fraction of E. coli. Maintenance and inner membrane-associated replication of plasmid RK2 were enhanced in a Dp knockout strain and inhibited in strains containing extra copies of the Dp gene or in membrane extracts to which a tagged form of Dp was added. Recently, the Dp was independently shown to help prevent overinitiation in E. coli and was termed Hda (S. Kato and T. Katayama, EMBO J. 20:4253-4262, 2001).

* Corresponding author. Mailing address: Department of Molecular Biology and Biochemistry, Wesleyan University, Middletown, CT 06459. Phone: (860) 685-2432. Fax: (860) 685-2141. E-mail: wfirshein{at}wesleyan.edu.

Journal of Bacteriology, March 2003, p. 1817-1824, Vol. 185, No. 6
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.6.1817-1824.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kongsuwan, K., Josh, P., Picault, M. J., Wijffels, G., Dalrymple, B. (2006). The Plasmid RK2 Replication Initiator Protein (TrfA) Binds to the Sliding Clamp {beta} Subunit of DNA Polymerase III: Implication for the Toxicity of a Peptide Derived from the Amino-Terminal Portion of 33-Kilodalton TrfA.. J. Bacteriol. 188: 5501-5509 [Abstract] [Full Text]  
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