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Overproduction of DnaA protein stimulates initiation of chromosome and minichromosome replication in Escherichia coli

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Summary

Increased synthesis of DnaA protein, obtained with plasmids carrying the dnaA gene controlled by the heat inducible λpL promoter, stimulated initiation of replication from oriC about threefold. The overinitiation was determined both as an increase in copy number of a minichromosome and as an increase in chromosomal gene dosage of oriC proximal DNA. The additional replication forks which were initiated on the chromosome did not lead to an overall increase in DNA content. DNA/DNA hybridization showed an amplification encompassing less than a few hundred kilobases on each side of oriC. Kinetic studies showed that the overinitiation occurred very rapidly after the induction, and that the initiation frequency then decreased to a near normal frequency per oriC. The results indicate that the DnaA protein is one important factor in regulation of initiation of DNA replication from oriC.

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Authors and Affiliations

  1. Department of Microbiology, The Technical University of Denmark, Building 221, DK-2800, Lyngby-Copenhagen, Denmark

    Tove Atlung, Anders Løbner-Olesen & Flemming G. Hansen

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  1. Tove Atlung
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  2. Anders Løbner-Olesen
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  3. Flemming G. Hansen
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Communicated by R. Devoret

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Atlung, T., Løbner-Olesen, A. & Hansen, F.G. Overproduction of DnaA protein stimulates initiation of chromosome and minichromosome replication in Escherichia coli . Mol Gen Genet 206, 51–59 (1987). https://doi.org/10.1007/BF00326535

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  • DOI: https://doi.org/10.1007/BF00326535

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