Home LiteratureArticle Details
PMID: 1729235 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

cysQ, a gene needed for cysteine synthesis in Escherichia coli K-12 only during aerobic growth.

Journal of bacteriology ·Vol. 174 ·No. 2 ·1992-01-00 ·Pages 415-25

Neuwald AF, Krishnan BR, Brikun I, Kulakauskas S, Suziedelis K, Tomcsanyi T, Leyh TS, Berg DE

Abstract

The initial steps in assimilation of sulfate during cysteine biosynthesis entail sulfate uptake and sulfate activation by formation of adenosine 5'-phosphosulfate, conversion to 3'-phosphoadenosine 5'-phosphosulfate, and reduction to sulfite. Mutations in a previously uncharacterized Escherichia coli gene, cysQ, which resulted in a requirement for sulfite or cysteine, were obtained by in vivo insertion of transposons Tn5tac1 and Tn5supF and by in vitro insertion of resistance gene cassettes. cysQ is at chromosomal position 95.7 min (kb 4517 to 4518) and is transcribed divergently from the adjacent cpdB gene. A Tn5tac1 insertion just inside the 3' end of cysQ, with its isopropyl-beta-D-thiogalactopyranoside-inducible tac promoter pointed toward the cysQ promoter, resulted in auxotrophy only when isopropyl-beta-D-thiogalactopyranoside was present; this conditional phenotype was ascribed to collision between converging RNA polymerases or interaction between complementary antisense and cysQ mRNAs. The auxotrophy caused by cysQ null mutations was leaky in some but not all E. coli strains and could be compensated by mutations in unlinked genes. cysQ mutants were prototrophic during anaerobic growth. Mutations in cysQ did not affect the rate of sulfate uptake or the activities of ATP sulfurylase and its protein activator, which together catalyze adenosine 5'-phosphosulfate synthesis. Some mutations that compensated for cysQ null alleles resulted in sulfate transport defects. cysQ is identical to a gene called amtA, which had been thought to be needed for ammonium transport. Computer analyses, detailed elsewhere, revealed significant amino acid sequence homology between cysQ and suhB of E. coli and the gene for mammalian inositol monophosphatase. Previous work had suggested that 3'-phosphoadenoside 5'-phosphosulfate is toxic if allowed to accumulate, and we propose that CysQ helps control the pool of 3'-phosphoadenoside 5'-phosphosulfate, or its use in sulfite synthesis.

Related Genes
MeSH Terms
Aerobiosis Alleles Amino Acid Sequence Ammonia/metabolism Base Sequence Cysteine/biosynthesis,genetics DNA, Bacterial/chemistry Escherichia coli/genetics,growth & development Gene Expression Regulation, Bacterial Genes, Bacterial/physiology Molecular Sequence Data Mutation Phenotype Transcription, Genetic
Chemicals
DNA, Bacterial Ammonia Cysteine
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Neuwald A F
Department of Molecular Microbiology, Washington University Medical School, St. Louis, Missouri 63110-1093.
Krishnan B R
Brikun I
Kulakauskas S
Suziedelis K
Tomcsanyi T
Leyh T S
Berg D E
References (47)
47 references, click to expand
  1. Thioredoxin or glutaredoxin in Escherichia coli is essential for sulfate reduction but not for deoxyribonucleotide synthesis.
    J Bacteriol. 1990 Apr;172(4):1923-9 PMID: 2180911
  2. Adenosine-5'-phosphosulfate kinase from Escherichia coli K12. Purification, characterization, and identification of a phosphorylated enzyme intermediate.
    J Biol Chem. 1989 Sep 5;264(25):15012-21 PMID: 2549047
  3. Sulfate and thiosulfate transport in Escherichia coli K-12: nucleotide sequence and expression of the cysTWAM gene cluster.
    J Bacteriol. 1990 Jun;172(6):3351-7 PMID: 2188958
  4. Cloning and expression of bovine brain inositol monophosphatase.
    J Biol Chem. 1990 Apr 15;265(11):5946-9 PMID: 1690719
  5. A mutation that enhances synthesis of sigma 32 and suppresses temperature-sensitive growth of the rpoH15 mutant of Escherichia coli.
    J Bacteriol. 1990 Apr;172(4):2124-30 PMID: 2138605
  6. Linkage map of Escherichia coli K-12, edition 8.
    Microbiol Rev. 1990 Jun;54(2):130-97 PMID: 2194094
  7. Sulfate and thiosulfate transport in Escherichia coli K-12: identification of a gene encoding a novel protein involved in thiosulfate binding.
    J Bacteriol. 1990 Jun;172(6):3358-66 PMID: 2188959
  8. Improved genetic selection for screening bacteriophage libraries by homologous recombination in vivo.
    Proc Natl Acad Sci U S A. 1990 Apr;87(8):3166-9 PMID: 2183225
  9. Identification of a sex-factor-affinity site in E. coli as gamma delta.
    Cold Spring Harb Symp Quant Biol. 1981;45 Pt 1:135-40 PMID: 6271456
  10. Polarity of Tn5 insertion mutations in Escherichia coli.
    J Bacteriol. 1980 May;142(2):439-46 PMID: 6247321
  11. Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements.
    Gene. 1982 Dec;20(2):305-16 PMID: 6299895
  12. Fine-structure deletion map and complementation analysis of the glnA-glnL-glnG region in Escherichia coli.
    J Bacteriol. 1982 Jun;150(3):1302-13 PMID: 6122676
  13. New M13 vectors for cloning.
    Methods Enzymol. 1983;101:20-78 PMID: 6310323
  14. Isolation of Escherichia coli mutants (cpdB) deficient in periplasmic 2':3'-cyclic phosphodiesterase and genetic mapping of the cpdB locus.
    J Gen Microbiol. 1980 Jul;119(1):31-4 PMID: 6251162
  15. Two separate genes involved in sulphate transport in Escherichia coli K12.
    J Gen Microbiol. 1983 Feb;129(2):357-8 PMID: 6341507
  16. Assimilatory sulfate reduction in Escherichia coli: identification of the alternate cofactor for adenosine 3'-phosphate 5'-phosphosulfate reductase as glutaredoxin.
    J Bacteriol. 1981 Jun;146(3):1059-66 PMID: 7016827
  17. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli.
    J Mol Biol. 1980 Apr;138(2):179-207 PMID: 6997493
  18. Cysteine auxotrophs of Salmonella typhimurium which grow without cysteine in a hydrogen/carbon dioxide atmosphere.
    J Gen Microbiol. 1979 Dec;115(2):513-6 PMID: 393803
  19. Deletions generated by the transposon Tn10 in the srl recA region of the Escherichia coli K-12 chromosome.
    Genetics. 1979 Oct;93(2):321-43 PMID: 395024
  20. Regulation of the assimilation of nitrogen compounds.
    Annu Rev Biochem. 1978;47:1127-62 PMID: 28074
  21. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  22. Sulfate-reducing pathway in Escherichia coli involving bound intermediates.
    J Bacteriol. 1976 Mar;125(3):923-33 PMID: 3497
  23. Appearance of double mutants in aged cultures of Salmonella typhimurium cysteine-requiring strains.
    Genetics. 1968 Aug;59(4):433-42 PMID: 4884730
  24. A binding site for sulfate and its relation to sulfate transport into Salmonella typhimurium.
    J Biol Chem. 1966 Sep 10;241(17):3962-9 PMID: 5331786
  25. CHARACTERIZATION OF A SULFATE- AND THIOSULFATE-TRANSPORTING SYSTEM IN SALMONELLA TYPHIMURIUM.
    J Biol Chem. 1964 Jul;239:2292-7 PMID: 14209960
  26. Acetylornithinase of Escherichia coli: partial purification and some properties.
    J Biol Chem. 1956 Jan;218(1):97-106 PMID: 13278318
  27. Studies on the mechanism of inhibition of Salmonella typhimurium by 1,2,4-triazole.
    J Biol Chem. 1975 Sep 25;250(18):7324-31 PMID: 1100624
  28. Sensitive mutants of bacteriophage lambda.
    Virology. 1961 May;14:22-32 PMID: 13690195
  29. The cysP promoter of Salmonella typhimurium: characterization of two binding sites for CysB protein, studies of in vivo transcription initiation, and demonstration of the anti-inducer effects of thiosulfate.
    J Bacteriol. 1991 Sep;173(18):5876-86 PMID: 1909324
  30. Transposon Tn5supF-based reverse genetic method for mutational analysis of Escherichia coli with DNAs cloned in lambda phage.
    J Bacteriol. 1991 Jan;173(2):896-9 PMID: 1846153
  31. Diverse proteins homologous to inositol monophosphatase.
    FEBS Lett. 1991 Dec 2;294(1-2):16-8 PMID: 1660408
  32. Efficient introduction of cloned mutant alleles into the Escherichia coli chromosome.
    J Bacteriol. 1991 Apr;173(8):2633-8 PMID: 1826503
  33. Ammonium transport in Escherichia coli: localization and nucleotide sequence of the amtA gene.
    J Gen Microbiol. 1991 Apr;137(4):983-9 PMID: 1856684
  34. PROSITE: a dictionary of sites and patterns in proteins.
    Nucleic Acids Res. 1991 Apr 25;19 Suppl:2241-5 PMID: 2041810
  35. Localization of the Escherichia coli amtA gene to 95.8 minutes.
    J Bacteriol. 1991 Mar;173(5):1572-3 PMID: 1999381
  36. Transcription and regulation of the cpdB gene in Escherichia coli K12 and Salmonella typhimurium LT2: evidence for modulation of constitutive promoters by cyclic AMP-CRP complex.
    Mol Gen Genet. 1990 Jun;222(1):161-5 PMID: 2172762
  37. The sulfate activation locus of Escherichia coli K12: cloning, genetic, and enzymatic characterization.
    J Biol Chem. 1988 Feb 15;263(5):2409-16 PMID: 2828368
  38. Cloning and characterization of the gene encoding inorganic pyrophosphatase of Escherichia coli K-12.
    J Bacteriol. 1988 Dec;170(12):5901-7 PMID: 2848015
  39. Isolation and sequence analysis of the gene (cpdB) encoding periplasmic 2',3'-cyclic phosphodiesterase.
    J Bacteriol. 1986 Mar;165(3):1002-10 PMID: 3005231
  40. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library.
    Cell. 1987 Jul 31;50(3):495-508 PMID: 3038334
  41. Novel regulatory mutants of the phosphate regulon in Escherichia coli K-12.
    J Mol Biol. 1986 Sep 5;191(1):39-58 PMID: 3540312
  42. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
  43. Tn5supF, a 264-base-pair transposon derived from Tn5 for insertion mutagenesis and sequencing DNAs cloned in phage lambda.
    Proc Natl Acad Sci U S A. 1989 Aug;86(15):5908-12 PMID: 2548192
  44. A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli.
    Microbiol Rev. 1989 Mar;53(1):1-24 PMID: 2540407
  45. Isolation of an ammonium or methylammonium ion transport mutant of Escherichia coli and complementation by the cloned gene.
    J Bacteriol. 1989 Feb;171(2):996-1001 PMID: 2536689
  46. Tn5tac1, a derivative of transposon Tn5 that generates conditional mutations.
    Proc Natl Acad Sci U S A. 1988 Sep;85(17):6468-72 PMID: 2842773
  47. Identification and cloning of genes involved in anaerobic sulfite reduction by Salmonella typhimurium.
    J Bacteriol. 1990 Jul;172(7):4100-2 PMID: 2163396
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
1992-01-00
Pages
415-25
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205732
Subset
IM
Grants
NIAID NIH HHS · 5 T32 AI07172 · United States
CSAT SAMHSA HHS · 5-TI-LM07049 · United States
NIGMS NIH HHS · GM-37138 · United States
Databases
GENBANK
M79367, M79368, M79369, M79370, M79371, M79372, M80795, M84980, Z11492, Z11493
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com