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

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Summary

A λ hybrid phage (λSda1), 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.

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Authors and Affiliations

  1. Centre de Recherche de Biochimie et de Génétique Cellulaires du C.N.R.S., 118 route de Narbonne, F-31062, Toulouse Cedex, France

    Olivier Fayer & Jean-Michel Louarn

  2. Department of Cellular, Viral and Molecular Biology, University of Utah Medical Center, 84132, Salt Lake City, UT, USA

    Olivier Fayer & Costa Georgopoulos

Authors
  1. Olivier Fayer
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  2. Jean-Michel Louarn
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  3. Costa Georgopoulos
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Communicated by G.R. Smith

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Fayer, O., Louarn, JM. & Georgopoulos, C. Suppression of the Escherichia coli dnaA46 mutation by amplification of the groES and groEL genes. Molec Gen Genet 202, 435–445 (1986). https://doi.org/10.1007/BF00333274

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  • DOI: https://doi.org/10.1007/BF00333274

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