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PMID: 3016662 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mechanism of autonomous control of the Escherichia coli F plasmid: different complexes of the initiator/repressor protein are bound to its operator and to an F plasmid replication origin.

Nucleic acids research ·Vol. 14 ·No. 14 ·1986-07-25 ·Pages 5693-711

Masson L, Ray DS

Abstract

E protein, the 29 kd product of the F plasmid repE gene, plays both positive and negative roles in the autoregulation of F replication. We have cloned and expressed the repE gene in an inducible ATG-fusion vector and have detected specific binding of E protein to the repE operator and to four 19-base pair direct repeats (incB) within the F plasmid replication origin ori2. Binding of E protein at the repE operator occurs with higher affinity than at ori2(incB) and gives almost complete protection to at least 30 base pairs, whereas binding of E protein to the direct repeats in the ori2 region shows an alternating pattern of enhanced and reduced sensitivity to DNAase cleavage consistent with a protein-induced folding of the DNA. These results provide direct biochemical support for a model of F plasmid replication in which the E protein serves both as an initiator of replication and as an autorepressor of its own synthesis.

MeSH Terms
Base Sequence Coliphages/genetics DNA Replication DNA Restriction Enzymes DNA, Viral/genetics,isolation & purification Escherichia coli/genetics F Factor Operon Peptide Initiation Factors/metabolism Promoter Regions, Genetic Repressor Proteins/metabolism Transcription Factors/metabolism
Chemicals
DNA, Viral Peptide Initiation Factors Repressor Proteins Transcription Factors DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Masson L
Ray D S
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36 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/14.14.5693
Published
1986-07-25
Pages
5693-711
Language
English
Region
England
NLM ID
0411011
PMCID
PMC311586
Subset
IM
Grants
NIAID NIH HHS · AI10752 · United States
NCI NIH HHS · CA16042 · United States
External Links
PubMed source DOI: 10.1093/nar/14.14.5693 PMC Full Text View journal details
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