The F plasmid centromere, sopC, is required for full repression of the sopAB operon.
The SopB protein of the F plasmid has a dual role in the partition of F plasmid copies to daughter cells prior to division. It binds to the sopC centromere site to form the partition complex needed for stabilizing the plasmid, and it interacts with SopA to repress transcription of the sopAB operon, thus preventing the destabilization that results from excess SopB. We have isolated sop mutants by screening for unstable inheritance of mutagenized mini-F DNA. Four of the mutants resulted from different missense mutations in sopB. All four were deficient, to varying degrees, in autoregulation of Sop protein synthesis. The mutant proteins showed diminished capacity for reducing the linking number of mini-F and for destabilizing a plasmid carrying sopC, indicating that reduced ability to form a normal complex with sopC might underlie the autoregulation defect. Repression of the transcription of a sop promoter- lacZ fusion by SopA and SopB was strongly enhanced in the presence of sopC, in cis or in trans, and the enhancement was reduced or nullified when wild-type sopB was replaced by the mutant sopB alleles. A single 43 bp unit of sopC was almost as effective as sopC itself in enhancing repression. The results show that sopC is necessary for full repression of the sop promoter. They thus indicate a previously unsuspected role for this centromere site, and suggest that autoregulation and partition might normally be coordinated processes.
Yates
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Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40536, USA.
Lane
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Biek
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DP
D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002503:Centromere; D017931:DNA Primers; D009154:Mutation; D009876:Operon; D010957:Plasmids / Q000235:genetics*; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D017931:DNA Primers; D012097:Repressor Proteins; C060972:SopB protein, Bacteria