Resolution of Holliday junctions in Escherichia coli: identification of the ruvC gene product as a 19-kilodalton protein

Author:

Sharples G J1,Lloyd R G1

Affiliation:

1. Department of Genetics, University of Nottingham, Queens Medical Centre, United Kingdom.

Abstract

The ruvC gene of Escherichia coli specifies a nuclease that resolves Holliday junction intermediates in genetic recombination (B. Connolly, C.A. Parsons, F.E. Benson, H.J. Dunderdale, G.J. Sharples, R.G. Lloyd, and S.C. West, Proc. Natl. Acad, Sci. USA 88:6063-6067, 1991). The gene was located between aspS and the ruvAB operon by DNA sequencing and deletion analysis of ruvC plasmids and was shown to encode a protein of 18,747 Da. Analysis of the DNA flanking ruvC indicated that the gene is transcribed independently of the LexA-regulated ruvAB operon and is not under direct SOS control. ruvC lies downstream of an open reading frame, orf-33, for a protein which migrates during sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a 33-kDa polypeptide. These two genes probably form an operon. However, expression of ruvC was found to be very poor relative to that of orf-33. A double ribosomal frameshift between these genes is proposed as a possible reason for the low level of RuvC. Two further open reading frames of unknown function were identified, one on either side of the orf-33-ruvC operon.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

Reference33 articles.

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2. Bachmann B. J. 1987. Derivations and genotypes of some mutant derivatives of Escherichia coli K-12 p. 1190-1219. In F. C. Neidhardt J. L. Ingraham K. B. Low B. Magasanik M. Schaechter and H. E. Umbarger (ed.) Escherichia coli and Salmonella typhimurium: cellular and molecular biology. American Society for Microbiology Washington D.C.

3. Regulation of two nested proteins from gene 49 (recombination endonuclease VII) and of a X RexA-like protein of bacteriophage T4;Barth K. A.;Genetics,1988

4. Nucleotide sequencing of the ruv region of Escherichia coli K-12 reveals a LexA regulated operon encoding two genes;Benson F. E.;Nucleic Acids Res.,1988

5. Resolution of Holliday junctions in vitro requires Escherichia coli ruvC gene product;Connolly B.;Proc. Natl. Acad. Sci. USA,1991

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