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• W1509665020 abstract "I described the co-association, in vitrointeraction, and epistatic relationship between the DNA replicationinitiator DnaA and the nucleoid-associated protein Rok in Bacillussubtilis. Using ChIP-seq, I characterized the genome-wideassociation of DnaA and its regulator YabA. I found that DnaA andYabA associated with over 30 chromosomal regions that were bound byRok, and association of DnaA and YabA with these regions wasdependent on Rok. The DNA-binding domain of DnaA was dispensablefor association of DnaA with these chromosomal regions. Thisindirect Rok-dependent association contrasted with the canonicalsequence-specific binding of DnaA to eight chromosomal regionscontaining clusters of DnaA boxes. I found that association of YabAwith Rok-dependent regions did not require DnaA, in contrast to theDnaA-dependent association of YabA with DnaA box cluster regions.Furthermore, I showed an in vitro interaction between DnaA and Rokusing purified proteins in an electrophoretic mobility shift assay.DnaA depended on Rok for association with a DNA probe,recapitulating the dependence observed at this chromosomal regionin vivo. DnaA and Rok are both transcription factors and regulatesome of the same genes. I analyzed global gene expression todetermine the genetic relationship between these twotranscriptional regulators. In general, rok was epistatic to dnaA;that is, the gene expression effects of DnaA depended on Rok,consistent with the ChIP-seq dependence. I investigated a potentialrole for Rok in regulating replication initiation, but I did notdetect a replication phenotype of a rok null mutant under variousconditions. I found that a rok null mutation did not detectablyaffect DnaA or YabA protein levels, and I showed that thestationary phase growth defect of this strain was dependent oncomK, a downstream target of Rok. Additionally, I determined that aputative regulator of replication initiation, the pyruvatedehydrogenase enzyme PdhC, affected replication via its metabolicfunction but was not a direct regulator of replicationinitation." @default.
• W1509665020 created "2016-06-24" @default.
• W1509665020 creator A5051760363 @default.
• W1509665020 date "2014-01-01" @default.
• W1509665020 modified "2023-09-23" @default.
• W1509665020 title "Genetic and biochemical interactions between the DNA replication initiator and a chromosome architecture protein in Bacillus subtilis" @default.
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