The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster

Tucker, Nicholas P., Hicks, Matthew G., Clarke, Thomas A. ORCID: https://orcid.org/0000-0002-6234-1914, Crack, Jason C., Chandra, Govind, Le Brun, Nick E. ORCID: https://orcid.org/0000-0001-9780-4061, Dixon, Ray and Hutchings, Matthew I. (2008) The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. PLoS One, 3 (11). ISSN 1932-6203

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Abstract

The regulatory protein NsrR, a member of the Rrf2 family of transcription repressors, is specifically dedicated to sensing nitric oxide (NO) in a variety of pathogenic and non-pathogenic bacteria. It has been proposed that NO directly modulates NsrR activity by interacting with a predicted [Fe-S] cluster in the NsrR protein, but no experimental evidence has been published to support this hypothesis. Here we report the purification of NsrR from the obligate aerobe Streptomyces coelicolor. We demonstrate using UV-visible, near UV CD and EPR spectroscopy that the protein contains an NO-sensitive [2Fe-2S] cluster when purified from E. coli. Upon exposure of NsrR to NO, the cluster is nitrosylated, which results in the loss of DNA binding activity as detected by bandshift assays. Removal of the [2Fe-2S] cluster to generate apo-NsrR also resulted in loss of DNA binding activity. This is the first demonstration that NsrR contains an NO-sensitive [2Fe-2S] cluster that is required for DNA binding activity.

Item Type:	Article
Additional Information:	© 2008 Tucker et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Faculty \ School:	Faculty of Science > School of Biological Sciences Faculty of Science > School of Chemistry (former - to 2024)
UEA Research Groups:	Faculty of Science > Research Groups > Molecular Microbiology Faculty of Science > Research Groups > Biophysical Chemistry (former - to 2017) Faculty of Science > Research Groups > Centre for Ocean and Atmospheric Sciences Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry Faculty of Science > Research Groups > Organisms and the Environment Faculty of Science > Research Groups > Chemistry of Life Processes Faculty of Science > Research Groups > Energy Materials Laboratory
Related URLs:	http://dx.doi.org/10.1371/journal.pone.0...
Depositing User:	Matthew Hutchings
Date Deposited:	29 Mar 2011 12:39
Last Modified:	25 Oct 2024 23:35
URI:	https://ueaeprints.uea.ac.uk/id/eprint/27317
DOI:	10.1371/journal.pone.0003623

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