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Nucleic Acids Research, 2002, Vol. 30, No. 5 1176-1181
© 2002 Oxford University Press

Directionality of {lambda} plasmid DNA replication carried out by the heritable replication complex

Sylwia Baranska1, Grazyna Konopa1 and Grzegorz Wegrzyn1,2,*

1Department of Molecular Biology, University of Gdaásk, Kßadki 24, 80-822 Gdaásk, Poland and 2Institute of Oceanology, Polish Academy of Sciences, ìw. Wojciecha 5, 81-347 Gdynia, Poland

There are two ‘pathways’ of replication of {lambda} plasmids in Escherichia coli. One pathway requires the assembly of a new replication complex before replication and the second pathway is based on the activity of the replication complex inherited by one of two daughter plasmid copies after a preceding replication round. Such a phenomenon was postulated to occur also in other replicons, including Saccharomyces cerevisiae autonomously replicating sequences. Here we investigated directionality of {lambda} plasmid replication carried out by the heritable and newly assembled replication complexes. Using two-dimensional agarose gel electrophoresis and electron microscopy we demonstrated that in both normal growth conditions and during the relaxed response to amino acid starvation (when only replication carried out by the heritable complex is possible), bidirectionally and undirectionally replicating plasmid molecules occurred in host cells in roughly equal proportions. The results are compatible with the hypothesis that both complexes (heritable and newly assembled) are equivalent.

* To whom correspondence should be addressed at: Department of Molecular Biology, University of Gdaásk, Kßadki 24, 80-822 Gdaásk, Poland. Tel: +48 58 346 3014; Fax: +48 58 301 0072; Email: wegrzyn{at}biotech.univ.gda.pl


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