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PMID: 3588285 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Cleavage of single-stranded DNA by plasmid pT181-encoded RepC protein.

Nucleic acids research ·Vol. 15 ·No. 10 ·1987-05-26 ·Pages 4085-97

Koepsel RR, Khan SA

Abstract

RepC protein encoded by plasmid pT181 has single-stranded endonuclease and topoisomerase-like activities. These activities may be involved in the initiation (and termination) of pT181 replication by a rolling circle mechanism. RepC protein cleaves the bottom strand of DNA within the origin of replication at a single, specific site when the DNA is in the supercoiled or linear (double or single-stranded) form. We have found that RepC protein will also cleave single-stranded DNA at sites other than the origin of replication. We have mapped the secondary cleavage sites on pT181 DNA. When the DNA is in the supercoiled, or linear, double-stranded form, only the primary site within the origin is cleaved. However, when the DNA is present in the single-stranded form, several strong and weak cleavage sites are observed. The DNA sequence at these cleavage sites shows a strong similarity with the primary cleavage site. The presence of Escherichia coli SSB protein inhibited cleavage at all of the secondary nick sites while the primary nick site remained susceptible to cleavage.

MeSH Terms
Bacterial Proteins/genetics,metabolism Base Sequence DNA Replication DNA, Bacterial/metabolism DNA, Single-Stranded/metabolism DNA-Binding Proteins/genetics,metabolism Plasmids Staphylococcus aureus/genetics,metabolism Substrate Specificity
Chemicals
Bacterial Proteins DNA, Bacterial DNA, Single-Stranded DNA-Binding Proteins RepC protein, Staphylococcus aureus
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Koepsel R R
Khan S A
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21 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.10.4085
Published
1987-05-26
Pages
4085-97
Language
English
Region
England
NLM ID
0411011
PMCID
PMC340833
Subset
IM
Grants
NIGMS NIH HHS · GM-3185 · United States
External Links
PubMed source DOI: 10.1093/nar/15.10.4085 PMC Full Text View journal details
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