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  • Title: Regulation of traJ transcription in the Salmonella virulence plasmid by strand-specific DNA adenine hemimethylation.
    Author: Camacho EM, Casadesús J.
    Journal: Mol Microbiol; 2005 Sep; 57(6):1700-18. PubMed ID: 16135235.
    Abstract:
    The traJ gene of the virulence plasmid of Salmonella enterica serovar Typhimurium (pSLT) encodes a transcriptional activator of the transfer operon. The leucine-responsive regulatory protein (Lrp) is an activator of traJ transcription. The upstream-activating-sequence of the pSLT traJ promoter contains two Lrp binding sites (LRP-1 and LRP-2), both necessary for transcriptional activation. The promoter-proximal site (LRP-2) contains a GATC site (GATC-II) whose methylation state affects Lrp binding: GATC-II methylation in both DNA strands decreases the affinity of Lrp for the LRP-2 site, while efficient Lrp binding occurs to a non-methylated GATC-II site. The effect of GATC-II hemimethylation on Lrp binding is strand-specific: methylation of the traJ non-coding strand permits formation of the major Lrp-DNA retardation complex, but methylation of the coding strand does not. This asymmetry supports a model in which passage of the replication fork may permit Lrp-mediated activation of conjugal transfer in one daughter plasmid molecule but not in the other. A remarkable trait of this regulatory design is that hemimethylation of a single GATC site can generate distinct epigenetic signals in otherwise identical plasmid DNA molecules.
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