The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster.
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] cl...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2008-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2577008?pdf=render |
| _version_ | 1818849432440930304 |
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| author | Nicholas P Tucker Matthew G Hicks Thomas A Clarke Jason C Crack Govind Chandra Nick E Le Brun Ray Dixon Matthew I Hutchings |
| author_facet | Nicholas P Tucker Matthew G Hicks Thomas A Clarke Jason C Crack Govind Chandra Nick E Le Brun Ray Dixon Matthew I Hutchings |
| author_sort | Nicholas P Tucker |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T06:33:09Z |
| format | Article |
| id | doaj.art-1ab9405ad66e4d42a2d2743213cd8894 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T06:33:09Z |
| publishDate | 2008-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-1ab9405ad66e4d42a2d2743213cd88942022-12-21T20:32:17ZengPublic Library of Science (PLoS)PLoS ONE1932-62032008-01-01311e362310.1371/journal.pone.0003623The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster.Nicholas P TuckerMatthew G HicksThomas A ClarkeJason C CrackGovind ChandraNick E Le BrunRay DixonMatthew I HutchingsThe 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.http://europepmc.org/articles/PMC2577008?pdf=render |
| spellingShingle | Nicholas P Tucker Matthew G Hicks Thomas A Clarke Jason C Crack Govind Chandra Nick E Le Brun Ray Dixon Matthew I Hutchings The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. PLoS ONE |
| title | The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. |
| title_full | The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. |
| title_fullStr | The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. |
| title_full_unstemmed | The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. |
| title_short | The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster. |
| title_sort | transcriptional repressor protein nsrr senses nitric oxide directly via a 2fe 2s cluster |
| url | http://europepmc.org/articles/PMC2577008?pdf=render |
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