Journal
JOURNAL OF BACTERIOLOGY
Volume 182, Issue 7, Pages 1995-2000Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JB.182.7.1995-2000.2000
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Funding
- NIGMS NIH HHS [R37 GM018277, R01 GM018277, F32 GM018277, GM18277] Funding Source: Medline
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Full activation of transcription of the araFGH promoter, p(FGH), requires both the catabolite activator protein (CAP) and AraC protein. At p(FGH), the binding site for CAP is centered at position -41.5, an essential binding site for AraC is centered at position -79.5, and a second, nonessential binding site is centered at position -154.5. In this work, we used the minimal promoter region required for in vivo activation of p(FGH) to examine the roles of CAP and AraC in stimulating formation of open complexes at p(FGH). Migration retardation assays of open complexes showed that RNA polymerase hinds exceptionally tightly to the AraC-CAP-p(FGH), complex and that the order of addition of proteins to the initiating complex is important. Similar assays with RNA polymerase containing truncated alpha subunits suggest that AraC interacts with the C-terminal domain of the alpha subunit. Finally, AraC protein also acts to prevent the improper binding of RNA polymerase at a pseudo promoter near the true p(FGH) promoter.
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