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PMID: 1986363 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Functional division and reconstruction of a plasmid replication origin: molecular dissection of the oriV of the broad-host-range plasmid RSF1010.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 1 ·1991-01-01 ·Pages 179-83

Honda Y, Sakai H, Hiasa H, Tanaka K, Komano T, Bagdasarian M

Abstract

Two single-stranded DNA initiation signals (designated ssi) present in the origin of vegetative DNA replication (oriV) of the broad-host-range plasmid RSF1010 are essential for the priming of replication of each complementary DNA strand of this plasmid in Escherichia coli. Each of the RSF1010 ssi signals, ssiA and ssiB, could be replaced by a primosome assembly site from plasmid pACY184 or from bacteriophage phi X174. In these chimeric origins, replication of the strand complementary to that containing the primosome assembly site was no longer dependent on the RSF1010 primase, protein RepB', but required the E. coli primase, DnaG. If both ssiA and ssiB sites of RSF1010 were replaced by primosome assembly sites, protein RepB' was no longer essential for the replication at this origin, whereas proteins RepA and RepC of RSF1010 were still required. These results strongly suggest that the two ssi sites and the RepB' protein actually direct the priming of DNA synthesis in the replication of RSF1010, and the proteins RepA and RepC are involved in the prepriming events--i.e., the opening of the DNA duplex at oriV. It is evident that the origin of RSF1010 can be separated into three functional domains and reconstructed by replacing the ssi sites with heterologous elements.

MeSH Terms
Base Sequence DNA Replication DNA, Bacterial/genetics Escherichia coli/genetics Models, Genetic Molecular Sequence Data Plasmids Restriction Mapping
Chemicals
DNA, Bacterial
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Honda Y
Department of Agricultural Chemistry, Kyoto University, Japan.
Sakai H
Hiasa H
Tanaka K
Komano T
Bagdasarian M
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31 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.1.179
Published
1991-01-01
Pages
179-83
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50773
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.88.1.179 PMC Full Text View journal details
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