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PMID: 3542972 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Repression and catabolite gene activation in the araBAD operon.

Journal of bacteriology ·Vol. 169 ·No. 2 ·1987-02-00 ·Pages 811-22

Lichenstein HS, Hamilton EP, Lee N

Abstract

Catabolite gene activation of the araBAD operon was examined by using catabolite gene activator protein (CAP) site deletion mutants. A high-affinity CAP-binding site between the divergently orientated araBAD and araC operons has been previously identified by DNase I footprinting techniques. Subsequent experiments disagreed as to whether this site is directly involved in stimulating araBAD expression. In this paper, we present data showing that deletions generated by in vitro mutagenesis of the CAP site led to a five- to sixfold reduction in single-copy araBAD promoter activity in vivo. We concluded that catabolite gene activation of araBAD involves this CAP site. The hypothesis that CAP stimulates the araBAD promoter primarily by relieving repression was then tested. The upstream operator araO2 was required for repression, but we observed that the magnitude of CAP stimulation was unaffected by the presence or absence of araO2. We concluded that CAP plays no role in relieving repression. Other experiments showed that when CAP binds it induces a bend in the ara DNA; similar bending has been reported upon CAP binding to lac DNA. This conformational change in the DNA may be essential to the mechanism of CAP activation.

MeSH Terms
Base Sequence Chromosome Deletion Escherichia coli/genetics Genes Genes, Bacterial Genes, Regulator Genetic Linkage Genotype Mutation Operon Promoter Regions, Genetic Receptors, Cyclic AMP/genetics
Chemicals
Receptors, Cyclic AMP
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lichenstein H S
Hamilton E P
Lee N
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61 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.169.2.811-822.1987
Published
1987-02-00
Pages
811-22
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC211852
Subset
IM
Grants
NIGMS NIH HHS · GM14652 · United States
External Links
PubMed source DOI: 10.1128/jb.169.2.811-822.1987 PMC Full Text View journal details
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