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Determination of VraR Binding Activity on fmtA Promoter
Author(s) -
Yang Zhifeng,
GolemiKotra Dasantila
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.769.2
Subject(s) - footprinting , binding site , transcription factor , penicillin binding proteins , transcription (linguistics) , electrophoretic mobility shift assay , chemistry , microbiology and biotechnology , peptidoglycan , promoter , biology , biochemistry , gene expression , gene , bacterial protein , linguistics , philosophy
In S. aureus , the VraSR two‐component system can sense cell wall stress and coordinate the response to antibiotics that inhibit cell wall peptidoglycan biosynthesis. fmtA encodes a low‐affinity penicillin binding protein and is part of the core cell wall stimulon. fmtA is described to be under the regulation of VraSR. In this study, we investigated binding of VraR to the fmtA promoter region. Two VraR binding regions in fmtA promoter have been found by EMSA and DNase I footprinting assays. The VraR binding motifs on these regions were predicted, and single or double nucleotide mutations were generated to investigate, the recognition or binding elements for VraR. We are using runoff in vitro transcription assays to investigate further the binding mode of VraR. The qRT‐PCR showed that the fmtA expression level was upregulated by VraS/VraR under oxacillin treatment. Our study suggests that VraR coordinates fmtA expression in vivo . Source of research support: CIHR Canadian Institutes of Health Research.