Differentiated, promoter-specific response of [4Fe-4S] NsrR DNA-binding to reaction with nitric oxide

Crack, Jason, Svistunenko, Dimitri A., Munnoch, John, Thomson, Andrew, Hutchings, Matthew and Le Brun, Nicolas ORCID: https://orcid.org/0000-0001-9780-4061 (2016) Differentiated, promoter-specific response of [4Fe-4S] NsrR DNA-binding to reaction with nitric oxide. Journal of Biological Chemistry, 291. pp. 8663-8672. ISSN 0021-9258

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Abstract

NsrR is an iron-sulfur cluster protein that regulates the nitric oxide (NO) stress response of many bacteria. NsrR from Streptomyces coelicolor regulates its own expression and that of only two other genes, hmpA1 and hmpA2, which encode HmpA enzymes predicted to detoxify NO. NsrR binds promoter DNA with high affinity only when coordinating a [4Fe-4S] cluster. Here we show that reaction of [4Fe-4S] NsrR with NO affects DNA-binding differently depending on the gene promoter. Binding to the hmpA2 promoter was abolished at ~2 NO per cluster, while for the hmpA1 and nsrR promoters, ~4 and ~8 NO molecules, respectively, were required to abolish DNA binding. Spectroscopic and kinetic studies of the NO reaction revealed a rapid, multi-phase, non-concerted process involving up to 8 – 10 NO molecules per cluster, leading to the formation of several iron-nitrosyl species. A distinct intermediate was observed at ~2 NO per cluster, along with two further intermediates at ~4 and ~6 NO. The NsrR nitrosylation reaction was not significantly affected by DNA-binding. These results show that NsrR regulates different promoters in response to different concentrations of NO. Spectroscopic evidence indicates that this is achieved by different NO-FeS complexes.

Item Type:	Article
Additional Information:	Final version free via Creative Commons CC-BY license
Uncontrolled Keywords:	iron,iron-sulfur,nitric oxide,regulator,dna-binding protein,spectroscopy
Faculty \ School:	Faculty of Science > School of Chemistry Faculty of Science > School of Biological Sciences
UEA Research Groups:	Faculty of Science > Research Groups > Biophysical Chemistry (former - to 2017) Faculty of Science > Research Groups > Molecular Microbiology 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
Depositing User:	Pure Connector
Date Deposited:	22 Mar 2016 09:31
Last Modified:	11 May 2023 13:30
URI:	https://ueaeprints.uea.ac.uk/id/eprint/57794
DOI:	10.1074/jbc.M115.693192

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