Activation by NarL at the Escherichia coli ogt promoter

Ruanto, Patcharawarin and Chismon, David L. and Hothersall, Joanne and Godfrey, Rita E. and Lee, David J. and Busby, Stephen J. W. and Browning, Douglas F. (2020) Activation by NarL at the Escherichia coli ogt promoter. Biochemical Journal, 477 (15). pp. 2807-2820. ISSN 1470-8728

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Abstract

The Escherichia coli NarX/NarL two-component response-regulator system regulates gene expression in response to nitrate ions and the NarL protein is a global transcription factor, which activates transcript initiation at many target promoters. One such target, the E. coli ogt promoter, which controls the expression of an O6-alkylguanine-DNA-alkyltransferase, is dependent on NarL binding to two DNA targets centred at positions −44.5 and −77.5 upstream from the transcript start. Here, we describe ogt promoter derivatives that can be activated solely by NarL binding either at position −44.5 or position −77.5. We show that NarL can also activate the ogt promoter when located at position −67.5. We present data to argue that NarL-dependent activation of transcript initiation at the ogt promoter results from a direct interaction between NarL and a determinant in the C-terminal domain of the RNA polymerase α subunit. Footprinting experiments show that, at the −44.5 promoter, NarL and the C-terminal domain of the RNA polymerase α subunit bind to opposite faces of promoter DNA, suggesting an unusual mechanism of transcription activation. Our work suggests new organisations for activator-dependent transcription at promoters and future applications for biotechnology.

Item Type: Article
Additional Information: ** Article version: VoR ** From Crossref via Jisc Publications Router ** History: epub 07-08-2020; issued 07-08-2020; ppub 14-08-2020. ** Licence for VoR version of this article starting on 14-07-2020: https://creativecommons.org/licenses/by/4.0/
Identification Number: https://doi.org/10.1042/bcj20200408
Date: 7 August 2020
Uncontrolled Keywords: Cell Biology, Biochemistry, Molecular Biology, molecular interactions, NarL, promoters, transcription activation
Subjects: C700 Molecular Biology, Biophysics and Biochemistry
Divisions: Faculty of Health, Education and Life Sciences > School of Health Sciences
REF UoA Output Collections > REF2021 UoA 03: Allied Health Professions, Dentistry, Nursing & Pharmacy
SWORD Depositor: JISC PubRouter
Depositing User: JISC PubRouter
Date Deposited: 19 Aug 2020 08:59
Last Modified: 18 Sep 2020 10:56
URI: http://www.open-access.bcu.ac.uk/id/eprint/9678

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