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The effects of an Escherichia coli dnaAts mutation on the replication of the plasmids colE1, pSC101, R100.1 and RTF-TC

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Summary

The rate of replication of the plasmids colE1, pSC101, R100.1 and pAR132 (an RTF-TC derivative of the drug resistance factor R100.1) has been investigated directly by DNA: DNA hybridization. These rates have been compared, in a dnaAts strain, to that of various markers of the host chromosome at permissive and non-permissive temperatures. Chromosome initiation in the dnaAts strain stops rapidly after a shift to the non-permissive temperature, but plasmids R100.1 and pAR132 do not seem to be affected directly and continue replication for some time. The colE1 replication rate undergoes a large increase after the temperature shift, followed by a rapid decrease to a very low level 25 min after the shift. In contrast pSC101 replication stops immediately after the shift. ColE1 is able to replicate in an integratively suppressed dnaAts strain at 42° C whereas pSC101 stops replication immediately under these conditions. We conclude that R100.1 and its derivative RTF-TC can replicate without a functional dnaA product; that colE1, while affected by a shift in temperature in a dnaAts strain, does not directly require dnaA; and that the plasmid pSC101 has an absolute requirement for dnaA. The absolute requirement of pSC101 for dnaA in the integratively suppressed Hfr strain provides a useful system for further investigations of the dnaA function.

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Authors and Affiliations

  1. Département de Biologie Moléculaire, 30, Quai Ernest-Ansermet, CH-1211, Genève 4, Switzerland

    J. Frey, M. Chandler & L. Caro

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  1. J. Frey
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  2. M. Chandler
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  3. L. Caro
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Communicated by W. Arber

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Frey, J., Chandler, M. & Caro, L. The effects of an Escherichia coli dnaAts mutation on the replication of the plasmids colE1, pSC101, R100.1 and RTF-TC. Molec. Gen. Genet. 174, 117–126 (1979). https://doi.org/10.1007/BF00268349

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  • DOI: https://doi.org/10.1007/BF00268349

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