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Abstract:
Regulators of DNA replication in bacteria are an attractive target for
new antibiotics, as not only is replication essential for cell
viability, but its underlying mechanisms also differ from those
operating in eukaryotes. The genetic information of most bacteria is
encoded on a single chromosome, but about 10% of species carry a split
genome spanning multiple chromosomes. The best studied bacterium in this
context is the human pathogen Vibrio cholerae, with a primary chromosome
(Chr1) of 3 M bps, and a secondary one (Chr2) of about 1 M bps.
Replication of Chr2 is under control of a unique mechanism, presenting a
potential target in the development of V. cholerae-specific antibiotics.
A common challenge in such endeavors is whether the effects of candidate
chemicals can be focused on specific mechanisms, such as DNA
replication. To test the specificity of antimicrobial substances
independent of other features of the V. cholerae cell for the
replication mechanism of the V. cholerae secondary chromosome, we
establish the replication machinery in the heterologous E. coli system.
We characterize an E. coli strain in which chromosomal replication is
driven by the replication origin of V. cholerae Chr2. Surprisingly, the
E. coli ori2 strain was not inhibited by vibrepin, previously found to
inhibit ori2-based replication.
