Partition of the Linear Plasmid N15: Interactions of N15 Partition Functions with the sop Locus of the F Plasmid

Author:

Ravin N.1,Lane D.2

Affiliation:

1. Bioengineering Centre, Russian Academy of Sciences, Moscow, 117312 Russia,1 and

2. Laboratoire de Microbiologie et Génétique Moléculaire, 31062 Toulouse, France2

Abstract

ABSTRACT A locus close to one end of the linear N15 prophage closely resembles the sop operon which governs partition of the F plasmid; the promoter region contains similar operator sites, and the two putative gene products have extensive amino acid identity with the SopA and -B proteins of F. Our aim was to ascertain whether the N15 sop homologue functions in partition, to identify the centromere site, and to examine possible interchangeability of function with the F Sop system. When expressed at a moderate level, N15 SopA and -B proteins partly stabilize mini-F which lacks its own sop operon but retains the sopC centromere. The stabilization does not depend on increased copy number. Likewise, an N15 mutant with most of its sop operon deleted is partly stabilized by F Sop proteins and fully stabilized by its own. Four inverted repeat sequences similar to those of sopC were located in N15. They are distant from the sop operon and from each other. Two of these were shown to stabilize a mini-F sop deletion mutant when N15 Sop proteins were provided. Provision of the SopA homologue to plasmids with a sopA deletion resulted in further destabilization of the plasmid. The N15 Sop proteins exert effective, but incomplete, repression at the F sop promoter. We conclude that the N15 sop locus determines stable inheritance of the prophage by using dispersed centromere sites. The SopB-centromere and SopA-operator interactions show partial functional overlap between N15 and F. SopA of each plasmid appears to interact with SopB of the other, but in a way that is detrimental to plasmid maintenance.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

Reference45 articles.

1. Two mini-F encoded proteins are essential for equipartition;Austin S.;Plasmid,1983

2. Bacterial plasmids that carry two functional centromere analogs are stable and are partitioned faithfully

3. Ausubel F. M. Brent R. Kingston R. E. Moore D. D. Seidman J. G. Smith J. A. Struhl K. Current protocols in molecular biology. 1989 Wiley-Interscience New York N.Y

4. A single 43-bp sopC repeat of plasmid mini-F is sufficient to allow assembly of a functional nucleoprotein partition complex;Biek D. P.;Proc. Natl. Acad. Sci. USA,1994

5. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli;Casadaban M. J.;J. Mol. Biol.,1980

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