PepA and ArgR do not regulate Cre recombination at the bacteriophage P1 loxP site

MacDonald, A.I., Lu, Y., Kilbride, E.A., Akopian, A. and Colloms, S.D. (2008) PepA and ArgR do not regulate Cre recombination at the bacteriophage P1 loxP site. Plasmid, 59(2), pp. 119-126. (doi:10.1016/j.plasmid.2007.12.001) (PMID:18226834) (PMCID:PMC2409434)

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Abstract

In the lysogenic state, bacteriophage P1 is maintained as a low copy-number circular plasmid. Site-specific recombination at loxP by the phage-encoded Cre protein keeps P1 monomeric, thus helping to ensure stable plasmid inheritance. Two Escherichia coli DNA-binding proteins, PepA and ArgR, were recently reported to be necessary for maintenance or establishment of P1 lysogeny. PepA and ArgR bind to regulatory DNA sequences upstream of the ColE1 cer recombination site to regulate site-specific recombination by the XerCD recombinases. This recombination keeps ColE1 in a monomeric state and helps to ensure stable plasmid maintenance. It has been suggested that ArgR and PepA play a similar role in P1 maintenance, regulating Cre recombination by binding to DNA sequences upstream of loxP. Here, we show that ArgR does not bind to its proposed binding site upstream of loxP, and that Cre recombination at loxP in its natural P1 context is not affected by PepA and ArgR in vitro. When sequences upstream of loxP were mutated to allow ArgR binding, PepA and ArgR still had no effect on Cre recombination. Our results demonstrate that PepA requires specific DNA sequences for binding, and that PepA and ArgR have no direct role in Cre recombination at P1 loxP.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:MacDonald, Dr Alasdair and Kilbride, Mrs Elizabeth and Colloms, Dr Sean
Authors: MacDonald, A.I., Lu, Y., Kilbride, E.A., Akopian, A., and Colloms, S.D.
Subjects:Q Science > QR Microbiology > QR355 Virology
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Plasmid
Publisher:Elsevier Inc.
ISSN:0147-619X
ISSN (Online):1095-9890
Published Online:28 January 2008
Copyright Holders:Copyright © 2007 Elsevier Inc.
First Published:First published in Plasmid 59(2):119-126
Publisher Policy:Reproduced under a creative commons license

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
252331Interactions between Proteins Bound at Distant Sites on DNA Molecules: Involvement in Site-Specific Recombination ...Sean CollomsWellcome Trust (WELLCOME)057651/Z/99/CH/RI MOLECULAR CELL & SYSTEMS BIOLOGY
252333Interactions between Proteins Bound at Distant Sites on DNA Molecules: Involvement in Site-Specific Recombination ...Sean CollomsWellcome Trust (WELLCOME)057651/Z/99/ARI MOLECULAR CELL & SYSTEMS BIOLOGY