title: Selenite and tellurite form mixed seleno- and tellurotrisulfides with CstR from Staphylococcus aureus.
abstract: Staphylococcus aureus CstR (CsoR-like sulfur transferase repressor) is a member of the CsoR family of transition metal sensing metalloregulatory proteins. Unlike CsoR, CstR does not form a stable complex with transition metals but instead reacts with sulfite to form a mixture of di- and trisulfide species, CstR2(RS-SR') and CstR2(RS-S-SR')n)n=1 or 2, respectively. Here, we investigate if CstR performs similar chemistry with related chalcogen oxyanions selenite and tellurite. In this work we show by high resolution tandem mass spectrometry that CstR is readily modified by selenite (SeO3(2-)) or tellurite (TeO3(2-)) to form a mixture of intersubunit disulfides and selenotrisulfides or tellurotrisulfides, respectively, between Cys31 and Cys60'. Analogous studies with S. aureus CsoR reveals no reaction with selenite and minimal reaction with tellurite. All cross-linked forms of CstR exhibit reduced DNA binding affinity. We show that Cys31 initiates the reaction with sulfite through the formation of S-sulfocysteine (RS-SO3(2-)) and Cys60 is required to fully derivatize CstR to CstR2(RS-SR') and CstR2(RS-S-SR'). The modification of Cys31 also drives an allosteric switch that negatively regulates DNA binding while derivatization of Cys60 alone has no effect on DNA binding. These results highlight the differences between CstRs and CsoRs in chemical reactivity and metal ion selectivity and establish Cys31 as the functionally important cysteine residue in CstRs.
authors: Department of Chemistry, Indiana University, Bloomington, IN 47405-7102, USA.
authors: Justin L
authors: JL
authors: Luebke
authors: Randy J
authors: RJ
authors: Arnold
authors: David P
authors: DP
authors: Giedroc
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism*; D002853:Chromatography, Liquid; D003432:Cross-Linking Reagents / Q000494:pharmacology; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D005454:Fluorescence Polarization; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D050505:Mutant Proteins / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D018038:Sodium Selenite / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D013211:Staphylococcus aureus / Q000187:drug effects / Q000378:metabolism*; D013440:Sulfides / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D013691:Tellurium / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D004269:DNA, Bacterial; D050505:Mutant Proteins; D010455:Peptides; D013440:Sulfides; D018038:Sodium Selenite; C026660:tellurous acid; D003545:Cysteine; D013691:Tellurium
references: Methods Enzymol. 1983;91:49-60
references: 6855597
references: Infect Disord Drug Targets. 2007 Jun;7(2):140-58
references: 17970225
references: J Biol Chem. 2012 May 18;287(21):17833-17847
references: 22451651
references: Microbiology (Reading). 2010 Jul;156(Pt 7):1993-2005
references: 20395270
references: Nature. 2010 Sep 23;467(7314):426-9
references: 20864996
references: ACS Chem Biol. 2010 Jan 15;5(1):47-62
references: 19957967
references: Biochemistry. 2012 Aug 28;51(34):6804-15
references: 22852582
references: J Biol Chem. 2011 Apr 15;286(15):13522-31
references: 21339296
references: Environ Sci Technol. 2007 Nov 1;41(21):7570-5
references: 18044543
references: Antioxid Redox Signal. 2011 Mar 15;14(6):1065-77
references: 20799881
references: Proc Natl Acad Sci U S A. 2002 May 14;99(10):6690-5
references: 11983871
references: Biochim Biophys Acta. 2005 May 25;1723(1-3):215-20
references: 15780970
references: FEMS Microbiol Rev. 2009 Jul;33(4):820-32
references: 19368559
references: J Bacteriol. 2004 Oct;186(20):6714-20
references: 15466022
references: PLoS Pathog. 2010 May 13;6(5):e1000894
references: 20485570
references: J Biol Chem. 2008 Aug 22;283(34):22890-4
references: 18593714
references: J Am Chem Soc. 2009 Dec 23;131(50):18044-5
references: 19928961
references: FEMS Microbiol Lett. 2004 Jun 15;235(2):357-62
references: 15183885
references: FEBS J. 2008 Jul;275(13):3352-61
references: 18494801
references: Mol Microbiol. 2009 Jul;73(2):194-211
references: 19508281
references: Curr Opin Chem Biol. 2008 Feb;12(1):18-24
references: 18282483
references: Nat Chem Biol. 2007 Jan;3(1):60-8
references: 17143269
references: Proc Natl Acad Sci U S A. 2007 Apr 3;104(14):5860-5
references: 17389395
references: N Engl J Med. 1998 Aug 20;339(8):520-32
references: 9709046
references: J Bacteriol. 2007 Nov;189(21):7911-9
references: 17720780
references: Acta Chem Scand. 1947;1(1):8-31
references: 20269018
references: J Biol Chem. 2009 Aug 28;284(35):23517-24
references: 19586910
references: Inorg Chem. 2009 Aug 17;48(16):7805-11
references: 19722686
references: J Am Chem Soc. 2005 Aug 24;127(33):11602-3
references: 16104727
references: J Biol Chem. 2012 Jun 15;287(25):21102-9
references: 22553203
references: Adv Microb Physiol. 2008;53:1-72
references: 17707143
references: J Biol Chem. 2011 Jul 22;286(29):26148-57
references: 21632538
references: Anal Chem. 2002 Sep 15;74(18):4602-10
references: 12349960
references: J Allergy Clin Immunol. 2007 Jul;120(1):13-22
references: 17606031
references: Proc Natl Acad Sci U S A. 2008 Sep 9;105(36):13586-91
references: 18757728
references: J Bacteriol. 2004 Mar;186(6):1579-90
references: 14996787
references: Annu Rev Biomed Eng. 2009;11:49-79
references: 19400705
references: Org Biomol Chem. 2010 Oct 7;8(19):4203-16
references: 20714663
references: Angew Chem Int Ed Engl. 2003 Oct 13;42(39):4742-58
references: 14562341
references: J Inorg Biochem. 2009 Mar;103(3):419-26
references: 19167089
references: Anal Biochem. 1996 Jun 1;237(2):260-73
references: 8660575
references: J Med Microbiol. 2007 Apr;56(Pt 4):454-458
references: 17374883
references: Nucleic Acids Res. 2012 May;40(9):4178-92
references: 22238377