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

Tucker, Nick and Hicks, Matthew G. and Clarke, Thomas A. and Crack, Jason C. and Chandra, Govinda and Le Brun, Nick E. and 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). e3623. ISSN 1932-6203 (https://doi.org/10.1371/journal.pone.0003623)

[thumbnail of journal.pone.0003623[1].pdf]
Preview
 PDF. Filename: journal.pone.0003623[1].pdf
Final Published Version
Download (573kB)| Preview

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.

ORCID iDs

Tucker, Nick ORCID logoORCID: https://orcid.org/0000-0002-6331-3704, Hicks, Matthew G., Clarke, Thomas A., Crack, Jason C., Chandra, Govinda, Le Brun, Nick E., Dixon, Ray and Hutchings, Matthew I.;
  • Item type:Article
    ID code:26108
    Dates:
    Date
    Event
    November 2008
    Published
    Notes:Strathprints' policy is to record up to 8 authors per publication, plus any additional authors based at the University of Strathclyde. More authors may be listed on the official publication than appear in the Strathprints' record.
    Subjects:Science > Chemistry
    Department:Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
    Depositing user: Strathprints Administrator
    Date deposited:24 Aug 2010 12:56
    Last modified:11 Nov 2024 09:32
    URI:https://strathprints.strath.ac.uk/id/eprint/26108
CORE (COnnecting REpositories)