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PMID: 3012269 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Suppression of the Escherichia coli dnaA46 mutation by amplification of the groES and groEL genes.

Molecular & general genetics : MGG ·Vol. 202 ·No. 3 ·1986-03-00 ·Pages 435-45

Fayet O, Louarn JM, Georgopoulos C

Abstract

A lambda hybrid phage (lambda Sda1), containing an 8.1 kb EcoRI DNA fragment from the Escherichia coli chromosome, was selected on the basis of its ability to suppress bacterial thermosensitivity caused by the dnaA46 mutation. We have shown that this suppression is due to a recA+-dependent amplification of the 8.1 kb fragment; consistent with this observation, cloning of the 8.1 kb fragment into a high copy number plasmid (pBR325) leads also to suppression of dnaA46. In the suppressed strains growing at high temperature, bidirectional replication starts in or near the oriC region and requires the presence of the DnaA polypeptide. These findings suggest that the overproduction of a gene product(s), encoded by the cloned 8.1 kb fragment, can restore dnaA-dependent initiation of replication at high temperature in the oriC region. Genetic mapping shows that the groES (mopB) and groEL (mopA) genes are located on the 8.1 kb suppressor fragment. Further analysis, including in vitro mutagenesis and subcloning, demonstrates that the amplification of the groES and groEL genes is both necessary and sufficient to suppress the temperature sensitive phenotype of the dnaA46 mutation.

MeSH Terms
Alleles Bacteriophage lambda/genetics Chromosome Mapping Chromosomes, Bacterial/physiology DNA Restriction Enzymes Escherichia coli/genetics Gene Amplification Genes, Bacterial Genotype Mutation Plasmids Species Specificity Suppression, Genetic
Chemicals
DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fayet O
Louarn J M
Georgopoulos C
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48 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00333274
Published
1986-03-00
Pages
435-45
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Grants
NIGMS NIH HHS · GM23917 · United States
External Links
PubMed source DOI: 10.1007/BF00333274
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