title: Staphylococcal NreB: an O(2)-sensing histidine protein kinase with an O(2)-labile iron-sulphur cluster of the FNR type.
abstract: The nreABC (nitrogen regulation) operon encodes a new staphylococcal two-component regulatory system that controls dissimilatory nitrate/nitrite reduction in response to oxygen. Unlike other two-component sensors NreB is a cytosolic protein with four N-terminal cysteine residues. It was shown that both the NreB-cysteine cluster and Fe ions are required for function. Isolated NreB was converted to the active form by incubation with cysteine desulphurase, ferrous ions and cysteine. This activation is typical for FeS-containing proteins and was reversed by oxygen. During reconstitution an absorption band at 420 nm and a yellow-brownish colour (typical for an FNR-type iron-sulphur cluster formation) developed. After alkylation of thiol groups in NreB and in the cysteine mutant NreB(C62S) almost no iron-sulphur cluster was incorporated; both findings corroborated the importance of the cysteine residues. Comparison of the kinase activity of (i). the reconstituted (ii). the unreconstituted, and (iii). the unreconstituted and deferrated NreB-His indicated that NreB kinase activity depended on iron availability and was greatly enhanced by reconstitution. NreB is the first direct oxygen-sensing protein described in staphylococci so far. Reconstituted NreB contains 4-8 acid-labile Fe and sulphide ions per NreB which is in agreement with the presence of 1-2 iron-sulphur [4Fe-4S](2+) clusters of the FNR-type. Unlike FNR, NreB does not act directly as transcriptional activator, but transfers the phosphoryl group to the response regulator NreC.
authors.affiliation: Mikrobielle Genetik, Universität Tübingen, D-72076 Tübingen, Germany.
authors.forename: Annegret
authors.initials: A
authors.lastname: Kamps
authors.forename: Stephanie
authors.initials: S
authors.lastname: Achebach
authors.forename: Iris
authors.initials: I
authors.lastname: Fedtke
authors.forename: Gottfried
authors.initials: G
authors.lastname: Unden
authors.forename: Friedrich
authors.initials: F
authors.lastname: Götz
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D000071677:Histidine Kinase; D007501:Iron / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism; D008954:Models, Biological; D009572:Nitrite Reductases / Q000378:metabolism; D009876:Operon; D010100:Oxygen / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013210:Staphylococcus / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D000255:Adenosine Triphosphate; D007501:Iron; D009572:Nitrite Reductases; D011494:Protein Kinases; D000071677:Histidine Kinase; D003545:Cysteine; D010100:Oxygen