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PMID: 2002497 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of the F plasmid traY promoter in Escherichia coli by host and plasmid factors.

Journal of molecular biology ·Vol. 218 ·No. 1 ·1991-03-05 ·Pages 119-28

Silverman PM, Wickersham E, Harris R

Abstract

F plasmid DNA transfer (tra) gene expression in Escherichia coli is regulated by chromosome- and F-encoded gene products. To study the relationship among these regulatory factors, we constructed low-copy plasmids containing a phi(traY'-'lacZ)hyb gene that couples beta-galactosidase and Lac permease synthesis to the F plasmid traY promoter. Wild-type transformants maintained high levels of beta-galactosidase over a broad range of culture densities. Primer extension analysis of tra mRNA from F'lac and phi(traY'-'lacZ)hyb strains indicated very similar, though not identical, transcription initiation sites. Moreover, phi(traY'-'lacZ)hyb gene expression required both TraJ and SfrA, as does tra gene expression in F+ strains. beta-Galactosidase activity was reduced approximately 30-fold in the absence of TraJ, which could be supplied in cis or in trans. In a two-plasmid system in which TraJ was supplied in trans by a lac-traJ operon fusion, phi(traY'-'lacZ)hyb expression was a linear, saturable function of traJ expression. Enzyme activity was reduced approximately tenfold in sfrA mutants. That reduction could not be attributed to an effect on the TraJ level. Several other cellular or environmental variables had only a modest effect on phi(traY'-'lacZ)hyb expression. Hyperexpression was observed at high cell density (twofold) and in anaerobic cultures (1.2- to 1.5-fold). In contrast, expression was reduced twofold in integration host factor mutants.

Related Genes
tra
MeSH Terms
Aerobiosis Anaerobiosis Base Sequence Chromosomes, Bacterial Escherichia coli/genetics,metabolism F Factor Gene Expression Regulation, Bacterial Genotype Molecular Sequence Data Plasmids Promoter Regions, Genetic RNA, Messenger/genetics Restriction Mapping Transformation, Bacterial
Chemicals
RNA, Messenger
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Silverman P M
Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Wickersham E
Harris R
Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
0022-2836
Published
1991-03-05
Pages
119-28
Language
English
Region
England
NLM ID
2985088R
Subset
IM
Grants
NIGMS NIH HHS · GM38657 · United States
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