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PMID: 2551682 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The Escherichia coli regulatory protein OxyR discriminates between methylated and unmethylated states of the phage Mu mom promoter.

The EMBO journal ·Vol. 8 ·No. 8 ·1989-08-00 ·Pages 2403-10

Bölker M, Kahmann R

Abstract

Expression of the phage Mu mom gene is transcriptionally regulated by DNA methylation. Three GATC sites upstream of the mom promoter have to be methylated by the Escherichia coli deoxyadenosine methylase (Dam) to allow initiation of transcription. An E. coli dam strain was mutagenized with Tn5 in an attempt to isolate mutants which allow mom gene expression. Three independent Tn5 mutants were isolated, each mapped to a gene at 89.6 min which we designate momR. The wildtype gene was cloned and sequenced, it encodes a protein of 305 amino acids. The protein belongs to a group of related bacterial activators recently identified as the LysR family (Henikoff et al., 1988). MomR protein was overproduced and purified. Expression of momR is autoregulated; MomR binds to a 43 bp region upstream of its coding sequence. In the mom promoter MomR protects a 43 bp region containing the three GATC sites. Specific binding to these sequences was observed only with unmethylated DNA. Fortuitously, we learned that MomR is identical to OxyR, a regulatory protein responding to oxidative stress. We discuss the implications of this control for Mu development.

MeSH Terms
Amino Acid Sequence Autoradiography Bacterial Proteins/genetics Bacteriophage mu/genetics Base Sequence DNA Transposable Elements DNA-Binding Proteins Escherichia coli/enzymology,genetics Escherichia coli Proteins Gene Expression Regulation Methylation Methyltransferases/metabolism Molecular Sequence Data Mutation Operon Promoter Regions, Genetic Repressor Proteins/genetics Restriction Mapping Site-Specific DNA-Methyltransferase (Adenine-Specific) Transcription Factors Transcription, Genetic
Chemicals
Bacterial Proteins DNA Transposable Elements DNA-Binding Proteins Escherichia coli Proteins Repressor Proteins Transcription Factors oxyR protein, E coli Methyltransferases Dam methyltransferase Site-Specific DNA-Methyltransferase (Adenine-Specific) dam protein, E coli
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bölker M
Institut für Genbiologische Forschung Berlin GmbH, FRG.
Kahmann R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1989-08-00
Pages
2403-10
Language
English
Region
England
NLM ID
8208664
PMCID
PMC401183
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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