Home LiteratureArticle Details
PMID: 9495743 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Genetic and functional analysis of the styrene catabolic cluster of Pseudomonas sp. strain Y2.

Journal of bacteriology ·Vol. 180 ·No. 5 ·1998-03-00 ·Pages 1063-71

Velasco A, Alonso S, García JL, Perera J, Díaz E

Abstract

The chromosomal region of Pseudomonas sp. strain Y2 involved in the conversion of styrene to phenylacetate (upper catabolic pathway) has been cloned and sequenced. Four catabolic genes, styABCD, and two regulatory genes, stySR, were identified. This gene cluster when transferred to Escherichia coli W confers to this phenylacetate-degrading host the ability to grow on styrene as the sole carbon and energy source. Genes styABCD are homologous to those encoding the styrene upper catabolic pathway in Pseudomonas fluorescens ST. Northern blot analyses have confirmed that genes styABCD constitute a transcription unit. The transcription start site of the sty operon was mapped 33 nucleotides upstream of the styA translational start codon. The styS and styR genes, which form an independent transcriptional unit, are located upstream of the styABCD operon, and their gene products show high similarity to members of the superfamily of two-component signal transduction systems. The styS gene product is homologous to histidine kinase proteins, whereas the styR gene product exhibits similarity at its N-terminal domain with cluster 1 of receiver modules and at its C terminus with the LuxR/FixJ family 3 of DNA-binding domains. Expression of the catabolic operon decreased significantly in the absence of the stySR genes and was restored when the stySR genes were provided in trans in the presence of styrene, suggesting that the stySR system behaves as a styrene-inducible positive regulator of the styABCD operon. Finally, a gene encoding a phenylacetyl-coenzyme A ligase that catalyzes the first step in the phenylacetate catabolism (styrene lower catabolic pathway) has been identified upstream of the styS gene. This activity was found to be induced in Pseudomonas sp. strain Y2 cells grown on styrene but not present in cells grown on glycerol. These results strongly suggest that the genes responsible for the complete mineralization of styrene are clustered in the chromosome of Pseudomonas sp. strain Y2.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/chemistry,genetics Base Composition Base Sequence Cloning, Molecular Coenzyme A Ligases/genetics,metabolism Gene Expression Regulation, Bacterial Genes, Bacterial Genes, Regulator Histidine Kinase Molecular Sequence Data Open Reading Frames Operon Phenylacetates/metabolism Protein Kinases/chemistry,genetics Pseudomonas/genetics,metabolism Sequence Alignment Sequence Analysis, DNA Styrene Styrenes/metabolism Transcription, Genetic Transformation, Bacterial
Chemicals
Bacterial Proteins Phenylacetates StyR protein, Pseudomonas Styrenes Styrene Protein Kinases Histidine Kinase Coenzyme A Ligases phenylacetate - CoA ligase phenylacetic acid
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Velasco A
Department of Molecular Microbiology, Centro de Investigaciones Biológicas, CSIC, Madrid, Spain.
Alonso S
García J L
Perera J
Díaz E
References (42)
42 references, click to expand
  1. Communication modules in bacterial signaling proteins.
    Annu Rev Genet. 1992;26:71-112 PMID: 1482126
  2. Metabolism of Styrene Oxide and 2-Phenylethanol in the Styrene-Degrading Xanthobacter Strain 124X.
    Appl Environ Microbiol. 1989 Nov;55(11):2850-5 PMID: 16348047
  3. Purification and characterization of a two-component monooxygenase that hydroxylates nitrilotriacetate from "Chelatobacter" strain ATCC 29600.
    J Bacteriol. 1992 Feb;174(4):1179-88 PMID: 1735711
  4. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  5. Bacterial degradation of styrene involving a novel flavin adenine dinucleotide-dependent styrene monooxygenase.
    Appl Environ Microbiol. 1990 May;56(5):1347-51 PMID: 2339888
  6. The Q-linker: a class of interdomain sequences found in bacterial multidomain regulatory proteins.
    Protein Eng. 1989 May;2(7):535-43 PMID: 2664763
  7. Prediction of the occurrence of the ADP-binding beta alpha beta-fold in proteins, using an amino acid sequence fingerprint.
    J Mol Biol. 1986 Jan 5;187(1):101-7 PMID: 3959077
  8. Evidence for two functional gal promoters in intact Escherichia coli cells.
    J Biol Chem. 1981 Nov 25;256(22):11905-10 PMID: 6271763
  9. Rapid similarity searches of nucleic acid and protein data banks.
    Proc Natl Acad Sci U S A. 1983 Feb;80(3):726-30 PMID: 6572363
  10. Possible regulatory role for nonaromatic carbon sources in styrene degradation by Pseudomonas putida CA-3.
    Appl Environ Microbiol. 1995 Feb;61(2):544-8 PMID: 7574594
  11. Cloning and analysis of structural genes from Streptomyces pristinaespiralis encoding enzymes involved in the conversion of pristinamycin IIB to pristinamycin IIA (PIIA): PIIA synthase and NADH:riboflavin 5'-phosphate oxidoreductase.
    J Bacteriol. 1995 Sep;177(18):5206-14 PMID: 7665509
  12. Response regulators of bacterial signal transduction systems: selective domain shuffling during evolution.
    J Mol Evol. 1995 Feb;40(2):136-54 PMID: 7699720
  13. Microbial metabolism and biotransformations of styrene.
    J Appl Bacteriol. 1994 Dec;77(6):597-606 PMID: 7822218
  14. Transcriptional control of the Pseudomonas putida TOL plasmid catabolic pathways.
    Mol Microbiol. 1993 Sep;9(5):923-9 PMID: 7934920
  15. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.
    Nucleic Acids Res. 1994 Nov 11;22(22):4673-80 PMID: 7984417
  16. Molecular characterization of 4-hydroxyphenylacetate 3-hydroxylase of Escherichia coli. A two-protein component enzyme.
    J Biol Chem. 1994 Sep 9;269(36):22823-9 PMID: 8077235
  17. Structural conservation in the CheY superfamily.
    Biochemistry. 1993 Nov 9;32(44):11741-53 PMID: 8218244
  18. Aldehyde dehydrogenases: widespread structural and functional diversity within a shared framework.
    Protein Sci. 1993 Nov;2(11):1890-900 PMID: 8268800
  19. Evolutionary divergence of pobA, the structural gene encoding p-hydroxybenzoate hydroxylase in an Acinetobacter calcoaceticus strain well-suited for genetic analysis.
    Gene. 1993 Mar 15;125(1):25-33 PMID: 8449410
  20. Cloning and characterization of styrene catabolism genes from Pseudomonas fluorescens ST.
    Appl Environ Microbiol. 1996 Jan;62(1):121-7 PMID: 8572689
  21. Structure of the Escherichia coli response regulator NarL.
    Biochemistry. 1996 Aug 27;35(34):11053-61 PMID: 8780507
  22. Signal transduction via the multi-step phosphorelay: not necessarily a road less traveled.
    Cell. 1996 Sep 20;86(6):845-8 PMID: 8808618
  23. Cloning and characterization of the genes encoding nitrilotriacetate monooxygenase of Chelatobacter heintzii ATCC 29600.
    J Bacteriol. 1996 Nov;178(21):6123-32 PMID: 8892809
  24. Styrene metabolism in Exophiala jeanselmei and involvement of a cytochrome P-450-dependent styrene monooxygenase.
    Appl Environ Microbiol. 1996 Apr;62(4):1471-4 PMID: 8919815
  25. Molecular cloning and expression in different microbes of the DNA encoding Pseudomonas putida U phenylacetyl-CoA ligase. Use of this gene to improve the rate of benzylpenicillin biosynthesis in Penicillium chrysogenum.
    J Biol Chem. 1996 Dec 27;271(52):33531-8 PMID: 8969218
  26. Cloning, sequencing, and analysis of a gene cluster from Chelatobacter heintzii ATCC 29600 encoding nitrilotriacetate monooxygenase and NADH:flavin mononucleotide oxidoreductase.
    J Bacteriol. 1997 Feb;179(4):1112-6 PMID: 9023192
  27. Identification and sequence analysis of two regulatory genes involved in anaerobic toluene metabolism by strain T1.
    Appl Environ Microbiol. 1997 Feb;63(2):652-60 PMID: 9023943
  28. A bacterial basic region leucine zipper histidine kinase regulating toluene degradation.
    Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1453-8 PMID: 9037074
  29. 2-phenylethylamine catabolism by Escherichia coli K-12: gene organization and expression.
    Microbiology. 1997 Feb;143 ( Pt 2):513-8 PMID: 9043126
  30. Identification and characterization of IS1381, a new insertion sequence in Streptococcus pneumoniae.
    J Bacteriol. 1997 Apr;179(7):2459-63 PMID: 9079939
  31. The first structure of an aldehyde dehydrogenase reveals novel interactions between NAD and the Rossmann fold.
    Nat Struct Biol. 1997 Apr;4(4):317-26 PMID: 9095201
  32. Characterization of the genes encoding a receptor-like histidine kinase and a cognate response regulator from a biphenyl/polychlorobiphenyl-degrading bacterium, Rhodococcus sp. strain M5.
    J Bacteriol. 1997 Apr;179(8):2772-6 PMID: 9098081
  33. Molecular characterization of PadA, a phenylacetaldehyde dehydrogenase from Escherichia coli.
    FEBS Lett. 1997 Apr 7;406(1-2):23-7 PMID: 9109378
  34. Identification of a novel positive regulator of the 4-hydroxyphenylacetate catabolic pathway of Escherichia coli.
    Biochem Biophys Res Commun. 1997 Mar 27;232(3):759-65 PMID: 9126350
  35. Molecular cloning and analysis of the genes encoding the 4-hydroxyphenylacetate hydroxylase from Klebsiella pneumoniae.
    Arch Microbiol. 1997 Feb-Mar;167(2-3):160-6 PMID: 9133323
  36. Sequencing and functional analysis of styrene catabolism genes from Pseudomonas fluorescens ST.
    Appl Environ Microbiol. 1997 Jun;63(6):2232-9 PMID: 9172343
  37. Modular multidomain phosphoryl transfer proteins of bacteria.
    Curr Opin Struct Biol. 1997 Jun;7(3):407-15 PMID: 9204284
  38. Region 2.5 of the Escherichia coli RNA polymerase sigma70 subunit is responsible for the recognition of the 'extended-10' motif at promoters.
    EMBO J. 1997 Jul 1;16(13):4034-40 PMID: 9233812
  39. PAS domain S-boxes in Archaea, Bacteria and sensors for oxygen and redox.
    Trends Biochem Sci. 1997 Sep;22(9):331-3 PMID: 9301332
  40. Purification and characterization of phenylacetaldehyde reductase from a styrene-assimilating Corynebacterium strain, ST-10.
    Appl Environ Microbiol. 1997 Oct;63(10):3783-8 PMID: 9327541
  41. Indigo formation by microorganisms expressing styrene monooxygenase activity.
    Appl Environ Microbiol. 1997 Nov;63(11):4287-91 PMID: 9361415
  42. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.180.5.1063-1071.1998
Published
1998-03-00
Pages
1063-71
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC106992
Subset
IM
Databases
GENBANK
AJ000330
External Links
PubMed source DOI: 10.1128/JB.180.5.1063-1071.1998 PMC Full Text View journal details
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: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2