Determinants of segregational stability of the linear plasmid-prophage N15 of Escherichia coli.
N15 is a bacteriophage of Escherichia coli that resembles lambda, but, unlike lambda, it lysogenizes as a linear plasmid. We show that stable maintenance of this unusual plasmid-prophage depends on the parA and parB genes, relatives of the partition genes of F and P1 plasmids. ParB of N15, like its F- and P1-encoded homologues, destabilizes plasmids carrying its target centromere, when present in excess. Within the genome of N15, we identified four unlinked, palindromic sequences that can promote the ParB-mediated destabilization of a moderate-copy vector in cis. They are distant from the parAB operon, unlike the centromeric sites of F and P1. Each of these palindromes could interact in vivo and in vitro with ParB. Each, when cloned separately, had properties characteristic of centromeric sites: exerted incompatibility against the N15 prophage and mini-N15 plasmids, and stabilized a mini-P1 plasmid depleted of its own partition genes when ParA and ParB of N15 were supplied. A pair of sites was more effective than a single site. Two of the centromeric sites are located in the proximity of promoters of phage genes, suggesting that, in addition to their function in partitioning of N15 prophage, they may control expression of N15 lytic functions.
Grigoriev
P S
PS
Department of Biophysics, State Technical University, 19251 St. Petersburg, Russia.
Lobocka
M B
MB
D001483:Base Sequence; D002503:Centromere / Q000235:genetics; D020090:Chromosome Segregation; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000821:virology*; D005814:Genes, Viral / Q000235:genetics; D008242:Lysogeny / Q000235:genetics; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*; D011533:Proviruses / Q000235:genetics*; D014169:Transformation, Bacterial; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D004279:DNA, Viral; D014764:Viral Proteins