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Journal of Bacteriology, January 2000, p. 477-487, Vol. 182, No. 2
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Role and Mechanism of Action of C · PvuII, a Regulatory Protein Conserved among Restriction-Modification Systems

Roy M. Vijesurier,1,dagger Leon Carlock,1 Robert M. Blumenthal,2,* and Joan C. Dunbar1

Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, Michigan 48201,1 and Department of Microbiology and Immunology, Medical College of Ohio, Toledo, Ohio 43614-58062

Received 29 April 1999/Accepted 27 October 1999

The PvuII restriction-modification system is a type II system, which means that its restriction endonuclease and modification methyltransferase are independently active proteins. The PvuII system is carried on a plasmid, and its movement into a new host cell is expected to be followed initially by expression of the methyltransferase gene alone so that the new host's DNA is protected before endonuclease activity appears. Previous studies have identified a regulatory gene (pvuIIC) between the divergently oriented genes for the restriction endonuclease (pvuIIR) and modification methyltransferase (pvuIIM), with pvuIIC in the same orientation as and partially overlapping pvuIIR. The product of pvuIIC, C · PvuII, was found to act in trans and to be required for expression of pvuIIR. In this study we demonstrate that premature expression of pvuIIC prevents establishment of the PvuII genes, consistent with the model that requiring C · PvuII for pvuIIR expression provides a timing delay essential for protection of the new host's DNA. We find that the opposing pvuIIC and pvuIIM transcripts overlap by over 60 nucleotides at their 5' ends, raising the possibility that their hybridization might play a regulatory role. We furthermore characterize the action of C · PvuII, demonstrating that it is a sequence-specific DNA-binding protein that binds to the pvuIIC promoter and stimulates transcription of both pvuIIC and pvuIIR into a polycistronic mRNA. The apparent location of C · PvuII binding, overlapping the -10 promoter hexamer and the pvuIICR transcriptional starting points, is highly unusual for transcriptional activators.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Medical College of Ohio, Toledo, OH 43614-5806. Phone: (419) 383-5422. Fax: (419) 383-3002. E-mail: rblumenthal{at}mco.edu.

dagger Present address: Department of Pediatrics, Children's Hospital of Michigan, Detroit, MI 48201.

Journal of Bacteriology, January 2000, p. 477-487, Vol. 182, No. 2
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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