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The multiple roles of CRP at the complex acs promoter depend on activation region 2 and IHF.

TitleThe multiple roles of CRP at the complex acs promoter depend on activation region 2 and IHF.
Publication TypeJournal Article
Year of Publication2007
AuthorsSclavi, B, Beatty, CM, Thach, DS, Fredericks, CE, Buckle, M, Wolfe, AJ
JournalMol Microbiol
Volume65
Issue2
Pagination425-40
Date Published2007 Jul
ISSN0950-382X
KeywordsAcetate-CoA Ligase, Base Sequence, Cyclic AMP Receptor Protein, DNA-Directed RNA Polymerases, Escherichia coli Proteins, Gene Expression Regulation, Bacterial, Integration Host Factors, Models, Biological, Promoter Regions, Genetic, Transcription Factors, Transcriptional Activation
Abstract

acs encodes a high-affinity enzyme that permits survival during carbon starvation. As befits a survival gene, its transcription is subject to complex regulation. Previously, we reported that cAMP receptor protein (CRP) activates acs transcription by binding tandem DNA sites located upstream of the major acsP2 promoter and that the nucleoid protein IHF (integration host factor) binds three specific sites located just upstream. In vivo, the sequence that includes these IHF sites exerts a positive effect on CRP-dependent transcription, while a construct containing only the most proximal site exhibits reduced transcription compared with the full-length promoter or with a construct lacking all three IHF sites. Here, we defined the minimal system required for this IHF-dependent inhibition, showing it requires the promoter-distal CRP site and an amino acid residue located within activation region 2 (AR2), a surface determinant of CRP that interacts with RNA polymerase (RNAP). Surprisingly, for a Class III promoter, disruption of AR2 caused significant changes in the activity and structure of both the full-length promoter and the construct with the single proximal IHF site. We propose that AR2, together with IHF, mediates formation of a multi-protein complex, in which RNAP is stabilized in an open complex that remains poised on the promoter ready to respond rapidly to environmental changes.

DOI10.1111/j.1365-2958.2007.05797.x
Alternate JournalMol. Microbiol.
PubMed ID17630973