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Microbiology 147 (2001), 1923-1928
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Microbiology (2001), 147, 1923-1928.
© 2001 Society for General Microbiology

Genetics and Molecular Biology

The double mechanism of incompatibility between {lambda} plasmids and Escherichia coli dnaA(ts) host cells

Monika Glinkowska1, Grazyna Konopa1, Alicja Wegrzyn2, Anna Herman-Antosiewicz1, Christoph Weigel3, Harald Seitz3, Walter Messer3 and Grzegorz Wegrzyn1,4

Department of Molecular Biology, University of Gdask, Kladki 24, 80-822 Gdask, Poland1
Laboratory of Molecular Biology (affiliated with the University of Gdask), Polish Academy of Sciences, Kladki 24, 80-822 Gdask, Poland2
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin-Dahlem, Germany3
Marine Biology Centre, Polish Academy of Sciences, w. Wojciecha 5, 81-347 Gdynia, Poland4

Author for correspondence: Grzegorz Wegrzyn. Tel: +48 58 346 3014. Fax: +48 58 301 0072. e-mail: wegrzyn{at}biotech.univ.gda.pl

For plasmids derived from bacteriophage {lambda}, the initiation of bidirectional DNA replication from ori{lambda} depends on the stimulation of transcription from the pR promoter by the host replication initiator protein DnaA. Certain Escherichia coli dnaA(ts) mutants cannot be transformed by wild-type {lambda} plasmids even at the temperature permissive to cell growth. This plasmid–host incompatibility appeared to be due to inefficient stimulation of transcription from the pR promoter by the mutant DnaA protein. This paper shows that there is a second mechanism for the incompatibility between {lambda} plasmids and dnaA(ts) hosts, exemplified in this study by the dnaA46 mutant. This is based on the competition between the {lambda} P protein and the host DnaA and DnaC proteins for DnaB helicase. Both mechanisms must be operative for the incompatibility.

Keywords: {lambda} plasmid replication, dnaA mutants, DnaA protein functions, transcriptional activation of origin, DnaB helicase

Abbreviations: aCT, autoclaved chlortetracycline




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