title: Genetic requirements for replication initiation of the staphylococcal multiresistance plasmid pSK41.
abstract: Replication of staphylococcal multiresistance plasmid pSK41 is initiated by binding of the replication initiator protein (Rep) to the Rep boxes, a series of four direct repeats located centrally within the rep gene. A Staphylococcus aureus strain was engineered to provide Rep in trans, allowing localization of the pSK41 origin of replication (oriV) to a 185 bp segment, which included the Rep boxes and a series of downstream direct repeats. Deletion analysis of individual Rep boxes revealed that all four Rep boxes are required for maximum origin activity, with the deletion of one or more Rep boxes having a significant effect on the proficiency of replication. However, a hierarchy of importance was identified among the Rep boxes, which appears to be mediated by the minor sequence variations that exist between them. DNA binding studies with truncated Rep proteins have enabled the DNA binding domain to be localized to the N-terminal 134 amino acids of the protein.
authors.affiliation: School of Biological Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Liu
authors.affiliation: School of Biological Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Kwong
authors.affiliation: School of Biological Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Cindy K
authors.initials: CK
authors.lastname: Pon
authors.affiliation: School of Biological Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Skurray
authors.affiliation: School of Biological Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Neville
authors.initials: N
authors.lastname: Firth
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication; D024901:Drug Resistance, Multiple, Bacterial; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D018741:Replication Origin; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D004265:DNA Helicases