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J Bacteriol. Mar 1993; 175(5): 1457–1466.
PMCID: PMC193233

Genetic structure and regulation of the cysG gene in Salmonella typhimurium.

Abstract

Siroheme, a cofactor of both sulfite and nitrite reductase in Salmonella typhimurium, requires the cysG gene for its synthesis. Three steps are required to synthesize siroheme from uroporphyrinogen III, the last common intermediate in the heme and siroheme pathways. All previously characterized cysG mutants were shown to be defective for the synthesis of cobalamin (B12), which shares a common precursor with siroheme. Since few cysG auxotrophs had been previously analyzed and since there is no evidence of siroheme mutants outside of the cysG region, we sought to expand the analysis of the region by isolating more mutations and studying the transcriptional regulation of the cysG gene using lacZ fusions. We isolated and analyzed 66 cysG auxotrophs. All were defective for both siroheme and cobalamin synthesis. Five exceptional mutants were partially defective for the synthesis of both and appear to be leaky. Complementation tests with tandem duplications suggest that the mutations causing the Cys auxotrophy affect only one cistron. The cysG gene is transcribed in a clockwise direction; this was demonstrated by a method that permits determining the orientation of two genes of unknown orientation provided their relative map order is known. The cysG gene was not part of the cysteine regulon, but had a substantial basal level of expression which was induced fivefold when cells were grown anaerobically on nitrite. Finally, we used Mud-generated duplications to genetically determine the organization of the cysG and nirB genes.

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  • Balch WE, Wolfe RS. New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressureized atmosphere. Appl Environ Microbiol. 1976 Dec;32(6):781–791. [PMC free article] [PubMed]
  • Benson NR, Goldman BS. Rapid mapping in Salmonella typhimurium with Mud-P22 prophages. J Bacteriol. 1992 Mar;174(5):1673–1681. [PMC free article] [PubMed]
  • Blanche F, Debussche L, Thibaut D, Crouzet J, Cameron B. Purification and characterization of S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase from Pseudomonas denitrificans. J Bacteriol. 1989 Aug;171(8):4222–4231. [PMC free article] [PubMed]
  • Bochner BR, Huang HC, Schieven GL, Ames BN. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. [PMC free article] [PubMed]
  • Casadaban MJ, Cohen SN. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. [PMC free article] [PubMed]
  • Castilho BA, Olfson P, Casadaban MJ. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. [PMC free article] [PubMed]
  • Cole JA, Coleman KJ, Compton BE, Kavanagh BM, Keevil CW. Nitrite and ammonia assimilation by anaerobic continuous cultures of Escherichia coli. J Gen Microbiol. 1974 Nov;85(1):11–22. [PubMed]
  • Cole JA, Newman BM, White P. Biochemical and genetic characterization of nirB mutants of Escherichia coli K 12 pleiotropically defective in nitrite and sulphite reduction. J Gen Microbiol. 1980 Oct;120(2):475–483. [PubMed]
  • Coleman KJ, Cornish-Bowden A, Cole JA. Purification and properties of nitrite reductase from Escherichia coli K12. Biochem J. 1978 Nov 1;175(2):483–493. [PMC free article] [PubMed]
  • Cox R, Charles HP. Porphyrin-accumulating mutants of Escherichia coli. J Bacteriol. 1973 Jan;113(1):122–132. [PMC free article] [PubMed]
  • DEMEREC M, GILLESPIE DH, MIZOBUCHI K. GENETIC STRUCTURE OF THE CYST REGION OF THE SALMONELLA GENOME. Genetics. 1963 Aug;48:997–1009. [PMC free article] [PubMed]
  • Escalante-Semerena JC, Roth JR. Regulation of cobalamin biosynthetic operons in Salmonella typhimurium. J Bacteriol. 1987 May;169(5):2251–2258. [PMC free article] [PubMed]
  • Hughes KT, Roth JR. Directed formation of deletions and duplications using Mud(Ap, lac). Genetics. 1985 Feb;109(2):263–282. [PMC free article] [PubMed]
  • Jayaraman PS, Peakman TC, Busby SJ, Quincey RV, Cole JA. Location and sequence of the promoter of the gene for the NADH-dependent nitrite reductase of Escherichia coli and its regulation by oxygen, the Fnr protein and nitrite. J Mol Biol. 1987 Aug 20;196(4):781–788. [PubMed]
  • Jeter RM, Olivera BM, Roth JR. Salmonella typhimurium synthesizes cobalamin (vitamin B12) de novo under anaerobic growth conditions. J Bacteriol. 1984 Jul;159(1):206–213. [PMC free article] [PubMed]
  • Kredich NM. Regulation of L-cysteine biosynthesis in Salmonella typhimurium. I. Effects of growth of varying sulfur sources and O-acetyl-L-serine on gene expression. J Biol Chem. 1971 Jun 10;246(11):3474–3484. [PubMed]
  • Kung HF, Spears C, Greene RC, Weissbach H. Regulation of the terminal reactions in methionine biosynthesis by vitamin B 12 and methionine. Arch Biochem Biophys. 1972 May;150(1):23–31. [PubMed]
  • Macdonald H, Cole J. Molecular cloning and functional analysis of the cysG and nirB genes of Escherichia coli K12, two closely-linked genes required for NADH-dependent nitrite reductase activity. Mol Gen Genet. 1985;200(2):328–334. [PubMed]
  • MacDonald H, Pope NR, Cole JA. Isolation, characterization and complementation analysis of nirB mutants of Escherichia coli deficient only in NADH-dependent nitrite reductase activity. J Gen Microbiol. 1985 Oct;131(10):2771–2782. [PubMed]
  • Maloy SR, Nunn WD. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. [PMC free article] [PubMed]
  • Murphy MJ, Siegel LM. Siroheme and sirohydrochlorin. The basis for a new type of porphyrin-related prosthetic group common to both assimilatory and dissimilatory sulfite reductases. J Biol Chem. 1973 Oct 10;248(19):6911–6919. [PubMed]
  • Murphy MJ, Siegel LM, Tove SR, Kamin H. Siroheme: a new prosthetic group participating in six-electron reduction reactions catalyzed by both sulfite and nitrite reductases. Proc Natl Acad Sci U S A. 1974 Mar;71(3):612–616. [PMC free article] [PubMed]
  • Ota N, Galsworthy PR, Pardee AB. Genetics of sulfate transport by Salmonella typhimurium. J Bacteriol. 1971 Mar;105(3):1053–1062. [PMC free article] [PubMed]
  • Peakman T, Busby S, Cole J. Transcriptional control of the cysG gene of Escherichia coli K-12 during aerobic and anaerobic growth. Eur J Biochem. 1990 Jul 31;191(2):325–331. [PubMed]
  • Peakman T, Crouzet J, Mayaux JF, Busby S, Mohan S, Harborne N, Wootton J, Nicolson R, Cole J. Nucleotide sequence, organisation and structural analysis of the products of genes in the nirB-cysG region of the Escherichia coli K-12 chromosome. Eur J Biochem. 1990 Jul 31;191(2):315–323. [PubMed]
  • Powell KA, Cox R, McConville M, Charles HP. Mutations affecting porphyrin biosynthesis in Escherichia coli. Enzyme. 1973;16(1):65–73. [PubMed]
  • Sanderson KE, Roth JR. Linkage map of Salmonella typhimurium, edition VII. Microbiol Rev. 1988 Dec;52(4):485–532. [PMC free article] [PubMed]
  • Murphy MJ, Siegel LM, Kamin H, Rosenthal D. Reduced nicotinamide adenine dinucleotide phosphate-sulfite reductase of enterobacteria. II. Identification of a new class of heme prosthetic group: an iron-tetrahydroporphyrin (isobacteriochlorin type) with eight carboxylic acid groups. J Biol Chem. 1973 Apr 25;248(8):2801–2814. [PubMed]
  • Stewart V. Nitrate respiration in relation to facultative metabolism in enterobacteria. Microbiol Rev. 1988 Jun;52(2):190–232. [PMC free article] [PubMed]
  • Strauch KL, Lenk JB, Gamble BL, Miller CG. Oxygen regulation in Salmonella typhimurium. J Bacteriol. 1985 Feb;161(2):673–680. [PMC free article] [PubMed]
  • VOGEL HJ, BONNER DM. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed]
  • Warren MJ, Roessner CA, Santander PJ, Scott AI. The Escherichia coli cysG gene encodes S-adenosylmethionine-dependent uroporphyrinogen III methylase. Biochem J. 1990 Feb 1;265(3):725–729. [PMC free article] [PubMed]
  • Warren MJ, Stolowich NJ, Santander PJ, Roessner CA, Sowa BA, Scott AI. Enzymatic synthesis of dihydrosirohydrochlorin (precorrin-2) and of a novel pyrrocorphin by uroporphyrinogen III methylase. FEBS Lett. 1990 Feb 12;261(1):76–80. [PubMed]
  • Wu JY, Siegel LM, Kredich NM. High-level expression of Escherichia coli NADPH-sulfite reductase: requirement for a cloned cysG plasmid to overcome limiting siroheme cofactor. J Bacteriol. 1991 Jan;173(1):325–333. [PMC free article] [PubMed]
  • Xu K, Delling J, Elliott T. The genes required for heme synthesis in Salmonella typhimurium include those encoding alternative functions for aerobic and anaerobic coproporphyrinogen oxidation. J Bacteriol. 1992 Jun;174(12):3953–3963. [PMC free article] [PubMed]

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