title: Evidence that the CysG protein catalyzes the first reaction specific to B12 synthesis in Salmonella typhimurium, insertion of cobalt.
abstract: The cysG gene of Salmonella typhimurium is involved in synthesis of both cobalamin (B12) and siroheme (a cofactor required for SO3(2-) and NO2(2-) reductases). The failure to reduce SO3(2-) leads to cysteine auxotrophy, for which the enzyme is named. Although Escherichia coli does not synthesize B12 de novo, it possesses a very similar CysG enzyme which has been shown to catalyze two methylations (uroporphyrinogen III to precorrin-2), ring oxidation (precorrin-2 to factor II), and iron insertion (factor II to siroheme). In S. typhimurium, precorrin-2 is a precursor of both siroheme and B12. All previously known Salmonella cysG mutants are defective in the synthesis of both siroheme and cobalamin. We describe two new classes of cysG mutants that cannot synthesize B12 but still make siroheme. For class I mutants, exogenous cobalt corrects the B12 defect but inhibits ability to make siroheme; B12 synthesis is inhibited by added iron. Class II mutants are unaffected by exogenous cobalt, but their B12 defect is corrected by derepression of the B12 biosynthetic genes (cob). We propose that all mutants are defective in insertion of cobalt into factor II and that the Salmonella CysG enzyme normally catalyzes this insertion-the first reaction dedicated to cobalamin synthesis. Although E. coli does not make B12, its CysG enzyme has been shown in vitro to insert cobalt into factor II and may have evolved to support B12 synthesis in some ancestor common to Salmonella species and E. coli.
authors.affiliation: Department of Biology, University of Utah, Salt Lake City 84112, USA.
authors.forename: T G
authors.initials: TG
authors.lastname: Fazzio
authors.forename: J R
authors.initials: JR
authors.lastname: Roth
mesh_terms: D002874:Chromosome Mapping; D003035:Cobalt / Q000378:metabolism / Q000494:pharmacology; D003470:Culture Media; D003545:Cysteine / Q000096:biosynthesis / Q000378:metabolism; D006418:Heme / Q000031:analogs & derivatives / Q000096:biosynthesis; D007501:Iron / Q000494:pharmacology; D008715:Methionine / Q000096:biosynthesis / Q000494:pharmacology; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D016296:Mutagenesis; D009876:Operon; D010084:Oxidation-Reduction; D010641:Phenotype; D017354:Point Mutation; D012486:Salmonella typhimurium / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D013447:Sulfites / Q000378:metabolism; D014805:Vitamin B 12 / Q000096:biosynthesis*
chemical_list: D003470:Culture Media; D013447:Sulfites; D003035:Cobalt; D006418:Heme; C009272:siroheme; D008715:Methionine; D007501:Iron; D008780:Methyltransferases; C060508:uroporphyrin-III C-methyltransferase; D003545:Cysteine; D014805:Vitamin B 12