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PMID: 8002608 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The leucine-responsive regulatory protein of Escherichia coli negatively regulates transcription of ompC and micF and positively regulates translation of ompF.

Journal of bacteriology ·Vol. 177 ·No. 1 ·1995-01-00 ·Pages 103-13

Ferrario M, Ernsting BR, Borst DW, Wiese DE, Blumenthal RM, Matthews RG

Abstract

The two major porins of Escherichia coli K-12 strains, OmpC and OmpF, are inversely regulated with respect to one another. The expression of OmpC and OmpF has been shown to be influenced by the leucine-responsive regulatory protein (Lrp): two-dimensional gel electrophoresis of proteins from strains with and strains without a functional Lrp protein revealed that OmpC expression is increased in an lrp strain, while OmpF expression is decreased. In agreement with these findings, we now present evidence that transcriptional (operon) fusions of lacZ+ to ompC and micF are negatively regulated by Lrp. Lrp binds specifically to the intergenic region between micF and ompC, as indicated by mobility shift assays and by DNase I footprinting. The expression of an ompF'-lacZ+ gene (translational) fusion is increased 3.7-fold in an lrp+ background compared with an lrp background, but expression of an ompF-lacZ+ operon fusion is not. Studies of in vivo expression of the outer membrane porins during growth on glucose minimal medium showed that the OmpF/OmpC ratio is higher in lrp+ strains than it is in isogenic lrp strains. The effect of Lrp was not seen in a strain containing a deletion of micF. Our studies suggest that the positive effect of Lrp on OmpF expression stems from a negative effect of Lrp on the expression of micF, an antisense RNA that inhibits ompF translation.

Related Genes
micF ompC ompF
MeSH Terms
Bacterial Outer Membrane Proteins/biosynthesis,genetics Bacterial Proteins/metabolism Base Sequence DNA, Bacterial/metabolism DNA-Binding Proteins/metabolism Escherichia coli/genetics Escherichia coli Proteins Gene Expression Regulation, Bacterial Leucine-Responsive Regulatory Protein Molecular Sequence Data Osmotic Pressure Plasmids/genetics Porins/biosynthesis Protein Binding Protein Biosynthesis Recombinant Fusion Proteins/biosynthesis Regulatory Sequences, Nucleic Acid/genetics Transcription Factors Transcription, Genetic beta-Galactosidase/biosynthesis,genetics
Chemicals
Bacterial Outer Membrane Proteins Bacterial Proteins DNA, Bacterial DNA-Binding Proteins Escherichia coli Proteins Lrp protein, E coli Porins Recombinant Fusion Proteins Transcription Factors Leucine-Responsive Regulatory Protein beta-Galactosidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ferrario M
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109-1055.
Ernsting B R
Borst D W
Wiese D E
Blumenthal R M
Matthews R G
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.177.1.103-113.1995
Published
1995-01-00
Pages
103-13
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC176562
Subset
IM
Grants
NIGMS NIH HHS · 2-T32-GM07315 · United States
External Links
PubMed source DOI: 10.1128/jb.177.1.103-113.1995 PMC Full Text View journal details
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