title: The interaction of RepC initiator with iterons in the replication of the broad host-range plasmid RSF1010.
abstract: The replication origin of the broad host-range plasmid RSF1010 contains 3.5 copies of a 20mer iteron sequence that bind specifically to the plasmid-encoded initiator, RepC. Here we demonstrated that even a single iteron was bent upon binding of RepC. Moreover, the bending angle seems to become larger along with the increment of the number of iterons. In a mutational analysis of the iteron sequence, we isolated seven kinds of base-substitution mutants of iterons, and estimated the replication activity of these mutants in vivo. We found that each of the subsections in the 20mer iteron sequence made a distinct contribution to the initiation of RSF1010 DNA replication. With the binding assay of RepC and mutated iterons in vitro, we found that the formation of a productive RepC-iteron complex was required for the initiation of plasmid DNA replication.
authors.affiliation: Department of Agricultural Chemistry, Kyoto University, Japan.
authors.forename: D M
authors.initials: DM
authors.lastname: Miao
authors.forename: H
authors.initials: H
authors.lastname: Sakai
authors.forename: S
authors.initials: S
authors.lastname: Okamoto
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
authors.forename: M
authors.initials: M
authors.lastname: Okuda
authors.forename: Y
authors.initials: Y
authors.lastname: Honda
authors.forename: T
authors.initials: T
authors.lastname: Komano
authors.forename: M
authors.initials: M
authors.lastname: Bagdasarian
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000096:biosynthesis* / Q000235:genetics*; D011485:Protein Binding; D018741:Replication Origin
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C043650:RepC protein, Staphylococcus aureus