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

Nick Tucker, Matthew G. Hicks, Thomas A. Clarke, Jason C. Crack, Govinda Chandra, Nick E. Le Brun, Ray Dixon, Matthew I. Hutchings

Research output: Contribution to journalArticle

98 Citations (Scopus)

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.
LanguageEnglish
Article numbere3623
Number of pages7
JournalPLoS One
Volume3
Issue number11
DOIs
Publication statusPublished - Nov 2008

Fingerprint

Repressor Proteins
nitric oxide
Nitric Oxide
DNA
Streptomyces coelicolor
aerobes
Proteins
regulatory proteins
Transcription
Escherichia coli
Purification
Paramagnetic resonance
spectroscopy
Assays
Spectrum Analysis
Bacteria
Demonstrations
proteins
transcription (genetics)
repressor proteins

Keywords

  • biochemistry studies
  • nucleic acids
  • purines
  • pyrimidines
  • physiology
  • biochemistry of bacteria

Cite this

Tucker, N., Hicks, M. G., Clarke, T. A., Crack, J. C., Chandra, G., Le Brun, N. E., ... Hutchings, M. I. (2008). The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. PLoS One, 3(11), [e3623]. https://doi.org/10.1371/journal.pone.0003623
Tucker, Nick ; Hicks, Matthew G. ; Clarke, Thomas A. ; Crack, Jason C. ; Chandra, Govinda ; Le Brun, Nick E. ; Dixon, Ray ; Hutchings, Matthew I. / The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. In: PLoS One. 2008 ; Vol. 3, No. 11.
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Tucker, N, Hicks, MG, Clarke, TA, Crack, JC, Chandra, G, Le Brun, NE, Dixon, R & Hutchings, MI 2008, 'The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster' PLoS One, vol. 3, no. 11, e3623. https://doi.org/10.1371/journal.pone.0003623

The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. / Tucker, Nick; Hicks, Matthew G.; Clarke, Thomas A.; Crack, Jason C.; Chandra, Govinda; Le Brun, Nick E.; Dixon, Ray; Hutchings, Matthew I.

In: PLoS One, Vol. 3, No. 11, e3623, 11.2008.

Research output: Contribution to journalArticle

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AU - Le Brun, Nick E.

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