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Activation and repression of a σN-dependent promoter naturally lacking upstream activation sequences
Umeå University, Faculty of Science and Technology, Department of Molecular Biology (Faculty of Science and Technology). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
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2009 (English)In: Molecular Microbiology, ISSN 0950-382X, E-ISSN 1365-2958, Vol. 73, no 3, 419-433 p.Article in journal (Refereed) Published
Abstract [en]

The Pseudomonas sp. strain ADP protein AtzR is a LysR-type transcriptional regulator required for activation of the atzDEF operon in response to nitrogen limitation and cyanuric acid. Transcription of atzR is directed by the σ(N)-dependent promoter PatzR, activated by NtrC and repressed by AtzR. Here we use in vivo and in vitro approaches to address the mechanisms of PatzR activation and repression. Activation by NtrC did not require any promoter sequences other than the sigma(N) recognition motif both in vivo and in vitro, suggesting that NtrC activates PatzR in an upstream activation sequences-independent fashion. Regarding AtzR-dependent autorepression, our in vitro transcription experiments show that the concentration of AtzR required for repression of the PatzR promoter in vitro correlates with AtzR affinity for its binding site. In addition, AtzR prevents transcription from PatzR when added to a preformed E-sigma(N)-PatzR closed complex, but isomerization to an open complex prevents repression. Gel mobility shift and DNase I footprint assays indicate that DNA-bound AtzR and E-σ(N) are mutually exclusive. Taken together, these results strongly support the notion that AtzR represses transcription from PatzR by competing with E-σ(N) for their overlapping binding sites. There are no previous reports of a similar mechanism for repression of σ(N)-dependent transcription.

Place, publisher, year, edition, pages
2009. Vol. 73, no 3, 419-433 p.
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Medical and Health Sciences
URN: urn:nbn:se:umu:diva-33334DOI: 10.1111/j.1365-2958.2009.06779.xPubMedID: 19570137OAI: diva2:311554
Available from: 2010-04-21 Created: 2010-04-21 Last updated: 2010-08-12Bibliographically approved

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Shingler, Victoria
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Department of Molecular Biology (Faculty of Science and Technology)Umeå Centre for Microbial Research (UCMR)
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