• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Nov 1995; 177(21): 6160–6169.
PMCID: PMC177456

A modified two-component regulatory system is involved in temperature-dependent biosynthesis of the Pseudomonas syringae phytotoxin coronatine.

Abstract

Biosynthesis of the phytotoxin coronatine (COR) in Pseudomonas syringae pv. glycinea PG4180 is regulated by temperature at the transcriptional level. A 3.4-kb DNA fragment from the COR biosynthetic gene cluster restored temperature-regulated phytotoxin production to Tn5 mutants defective in COR production. Nucleotide sequence analysis of this fragment revealed three genes, corS, corP, and corR, which encode a modified two-component regulatory system consisting of one sensor protein, CorS, and two response regulator proteins, CorP and CorR. Although only one response regulator, CorR, had a DNA-binding domain, the phosphate-receiving domains of both response regulator proteins were highly conserved. Transcriptional fusions of the corP and corR promoters to a promoterless glucuronidase gene (uidA) indicated that these two genes are expressed constitutively at 18 and 28 degrees C. In contrast, a corS::uidA fusion exhibited the temperature dependence previously observed for COR biosynthetic promoters and exhibited maximal transcriptional activity at 18 degrees C and low activity at 28 degrees C. Furthermore, glucuronidase activity for corS::uidA was decreased in corP, corR, and corS mutants relative to the levels observed for PG4180(corS::uidA). This difference was not observed for corP::uidA and corR::uidA transcriptional fusions since expression of these fusions remained low and constitutive regardless of the genetic background. The three regulatory genes functioned in a P. syringae strain lacking the COR gene cluster to achieve temperature-dependent activation of an introduced COR biosynthetic promoter, indicating that this triad of genes is the primary control for COR biosynthesis and responsible for thermoregulation. Our data suggest that the modified two-component regulatory system described in this study might transduce and amplify a temperature signal which results in transcriptional activation of COR biosynthetic genes.

Full Text

The Full Text of this article is available as a PDF (801K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aricò B, Scarlato V, Monack DM, Falkow S, Rappuoli R. Structural and genetic analysis of the bvg locus in Bordetella species. Mol Microbiol. 1991 Oct;5(10):2481–2491. [PubMed]
  • Bender CL, Cooksey DA. Indigenous plasmids in Pseudomonas syringae pv. tomato: conjugative transfer and role in copper resistance. J Bacteriol. 1986 Feb;165(2):534–541. [PMC free article] [PubMed]
  • Bender CL, Liyanage H, Palmer D, Ullrich M, Young S, Mitchell R. Characterization of the genes controlling the biosynthesis of the polyketide phytotoxin coronatine including conjugation between coronafacic and coronamic acid. Gene. 1993 Oct 29;133(1):31–38. [PubMed]
  • Bender CL, Malvick DK, Mitchell RE. Plasmid-mediated production of the phytotoxin coronatine in Pseudomonas syringae pv. tomato. J Bacteriol. 1989 Feb;171(2):807–812. [PMC free article] [PubMed]
  • Bender CL, Young SA, Mitchell RE. Conservation of Plasmid DNA Sequences in Coronatine-Producing Pathovars of Pseudomonas syringae. Appl Environ Microbiol. 1991 Apr;57(4):993–999. [PMC free article] [PubMed]
  • Bishop RE, Weiner JH. Overproduction, solubilization, purification and DNA-binding properties of AmpR from Citrobacter freundii. Eur J Biochem. 1993 Apr 1;213(1):405–412. [PubMed]
  • Burbulys D, Trach KA, Hoch JA. Initiation of sporulation in B. subtilis is controlled by a multicomponent phosphorelay. Cell. 1991 Feb 8;64(3):545–552. [PubMed]
  • Cooper VJ, Salmond GP. Molecular analysis of the major cellulase (CelV) of Erwinia carotovora: evidence for an evolutionary "mix-and-match" of enzyme domains. Mol Gen Genet. 1993 Nov;241(3-4):341–350. [PubMed]
  • Deretic V, Dikshit R, Konyecsni WM, Chakrabarty AM, Misra TK. The algR gene, which regulates mucoidy in Pseudomonas aeruginosa, belongs to a class of environmentally responsive genes. J Bacteriol. 1989 Mar;171(3):1278–1283. [PMC free article] [PubMed]
  • Ferguson IB, Mitchell RE. Stimulation of ethylene production in bean leaf discs by the pseudomonad phytotoxin coronatine. Plant Physiol. 1985 Apr;77(4):969–973. [PMC free article] [PubMed]
  • Feys BJF, Benedetti CE, Penfold CN, Turner JG. Arabidopsis Mutants Selected for Resistance to the Phytotoxin Coronatine Are Male Sterile, Insensitive to Methyl Jasmonate, and Resistant to a Bacterial Pathogen. Plant Cell. 1994 May;6(5):751–759. [PMC free article] [PubMed]
  • Figurski DH, Helinski DR. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. [PMC free article] [PubMed]
  • Gunsalus RP, Kalman LV, Stewart RR. Nucleotide sequence of the narL gene that is involved in global regulation of nitrate controlled respiratory genes of Escherichia coli. Nucleic Acids Res. 1989 Mar 11;17(5):1965–1975. [PMC free article] [PubMed]
  • Hale TL. Genetic basis of virulence in Shigella species. Microbiol Rev. 1991 Jun;55(2):206–224. [PMC free article] [PubMed]
  • Hrabak EM, Willis DK. The lemA gene required for pathogenicity of Pseudomonas syringae pv. syringae on bean is a member of a family of two-component regulators. J Bacteriol. 1992 May;174(9):3011–3020. [PMC free article] [PubMed]
  • Huang J, Carney BF, Denny TP, Weissinger AK, Schell MA. A complex network regulates expression of eps and other virulence genes of Pseudomonas solanacearum. J Bacteriol. 1995 Mar;177(5):1259–1267. [PMC free article] [PubMed]
  • Huang J, Denny TP, Schell MA. vsrB, a regulator of virulence genes of Pseudomonas solanacearum, is homologous to sensors of the two-component regulator family. J Bacteriol. 1993 Oct;175(19):6169–6178. [PMC free article] [PubMed]
  • Hugouvieux-Cotte-Pattat N, Dominguez H, Robert-Baudouy J. Environmental conditions affect transcription of the pectinase genes of Erwinia chrysanthemi 3937. J Bacteriol. 1992 Dec;174(23):7807–7818. [PMC free article] [PubMed]
  • Island MD, Wei BY, Kadner RJ. Structure and function of the uhp genes for the sugar phosphate transport system in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1992 May;174(9):2754–2762. [PMC free article] [PubMed]
  • Iuchi S, Matsuda Z, Fujiwara T, Lin EC. The arcB gene of Escherichia coli encodes a sensor-regulator protein for anaerobic repression of the arc modulon. Mol Microbiol. 1990 May;4(5):715–727. [PubMed]
  • Jin S, Song YN, Deng WY, Gordon MP, Nester EW. The regulatory VirA protein of Agrobacterium tumefaciens does not function at elevated temperatures. J Bacteriol. 1993 Nov;175(21):6830–6835. [PMC free article] [PubMed]
  • Kenyon JS, Turner JG. The Stimulation of Ethylene Synthesis in Nicotiana tabacum Leaves by the Phytotoxin Coronatine. Plant Physiol. 1992 Sep;100(1):219–224. [PMC free article] [PubMed]
  • KING EO, WARD MK, RANEY DE. Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med. 1954 Aug;44(2):301–307. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Liyanage H, Penfold C, Turner J, Bender CL. Sequence, expression and transcriptional analysis of the coronafacate ligase-encoding gene required for coronatine biosynthesis by Pseudomonas syringae. Gene. 1995 Feb 3;153(1):17–23. [PubMed]
  • Matsumura P, Rydel JJ, Linzmeier R, Vacante D. Overexpression and sequence of the Escherichia coli cheY gene and biochemical activities of the CheY protein. J Bacteriol. 1984 Oct;160(1):36–41. [PMC free article] [PubMed]
  • Mekalanos JJ. Environmental signals controlling expression of virulence determinants in bacteria. J Bacteriol. 1992 Jan;174(1):1–7. [PMC free article] [PubMed]
  • Melton AR, Weiss AA. Environmental regulation of expression of virulence determinants in Bordetella pertussis. J Bacteriol. 1989 Nov;171(11):6206–6212. [PMC free article] [PubMed]
  • Msadek T, Kunst F, Henner D, Klier A, Rapoport G, Dedonder R. Signal transduction pathway controlling synthesis of a class of degradative enzymes in Bacillus subtilis: expression of the regulatory genes and analysis of mutations in degS and degU. J Bacteriol. 1990 Feb;172(2):824–834. [PMC free article] [PubMed]
  • Murillo J, Shen H, Gerhold D, Sharma A, Cooksey DA, Keen NT. Characterization of pPT23B, the plasmid involved in syringolide production by Pseudomonas syringae pv. tomato PT23. Plasmid. 1994 May;31(3):275–287. [PubMed]
  • Palmer DA, Bender CL. Effects of Environmental and Nutritional Factors on Production of the Polyketide Phytotoxin Coronatine by Pseudomonas syringae pv. Glycinea. Appl Environ Microbiol. 1993 May;59(5):1619–1626. [PMC free article] [PubMed]
  • Pao GM, Tam R, Lipschitz LS, Saier MH., Jr Response regulators: structure, function and evolution. Res Microbiol. 1994 Jun-Aug;145(5-6):356–362. [PubMed]
  • Parkinson JS, Kofoid EC. Communication modules in bacterial signaling proteins. Annu Rev Genet. 1992;26:71–112. [PubMed]
  • Parsek MR, Shinabarger DL, Rothmel RK, Chakrabarty AM. Roles of CatR and cis,cis-muconate in activation of the catBC operon, which is involved in benzoate degradation in Pseudomonas putida. J Bacteriol. 1992 Dec;174(23):7798–7806. [PMC free article] [PubMed]
  • Reitzer LJ, Magasanik B. Isolation of the nitrogen assimilation regulator NR(I), the product of the glnG gene of Escherichia coli. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5554–5558. [PMC free article] [PubMed]
  • Rowley KB, Clements DE, Mandel M, Humphreys T, Patil SS. Multiple copies of a DNA sequence from Pseudomonas syringae pathovar phaseolicola abolish thermoregulation of phaseolotoxin production. Mol Microbiol. 1993 May;8(3):625–635. [PubMed]
  • Schmitt MP, Holmes RK. Cloning, sequence, and footprint analysis of two promoter/operators from Corynebacterium diphtheriae that are regulated by the diphtheria toxin repressor (DtxR) and iron. J Bacteriol. 1994 Feb;176(4):1141–1149. [PMC free article] [PubMed]
  • Schröder I, Wolin CD, Cavicchioli R, Gunsalus RP. Phosphorylation and dephosphorylation of the NarQ, NarX, and NarL proteins of the nitrate-dependent two-component regulatory system of Escherichia coli. J Bacteriol. 1994 Aug;176(16):4985–4992. [PMC free article] [PubMed]
  • Sundin GW, Bender CL. Ecological and genetic analysis of copper and streptomycin resistance in Pseudomonas syringae pv. syringae. Appl Environ Microbiol. 1993 Apr;59(4):1018–1024. [PMC free article] [PubMed]
  • Stock JB, Ninfa AJ, Stock AM. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. [PMC free article] [PubMed]
  • Stout V, Gottesman S. RcsB and RcsC: a two-component regulator of capsule synthesis in Escherichia coli. J Bacteriol. 1990 Feb;172(2):659–669. [PMC free article] [PubMed]
  • Tobe T, Yoshikawa M, Sasakawa C. Thermoregulation of virB transcription in Shigella flexneri by sensing of changes in local DNA superhelicity. J Bacteriol. 1995 Feb;177(4):1094–1097. [PMC free article] [PubMed]
  • Ullrich M, Bender CL. The biosynthetic gene cluster for coronamic acid, an ethylcyclopropyl amino acid, contains genes homologous to amino acid-activating enzymes and thioesterases. J Bacteriol. 1994 Dec;176(24):7574–7586. [PMC free article] [PubMed]
  • Ullrich M, Guenzi AC, Mitchell RE, Bender CL. Cloning and expression of genes required for coronamic Acid (2-ethyl-1-aminocyclopropane 1-carboxylic Acid), an intermediate in the biosynthesis of the phytotoxin coronatine. Appl Environ Microbiol. 1994 Aug;60(8):2890–2897. [PMC free article] [PubMed]
  • Van den Eede G, Deblaere R, Goethals K, Van Montagu M, Holsters M. Broad host range and promoter selection vectors for bacteria that interact with plants. Mol Plant Microbe Interact. 1992 May-Jun;5(3):228–234. [PubMed]
  • Wanner BL. Is cross regulation by phosphorylation of two-component response regulator proteins important in bacteria? J Bacteriol. 1992 Apr;174(7):2053–2058. [PMC free article] [PubMed]
  • Weiler EW, Kutchan TM, Gorba T, Brodschelm W, Niesel U, Bublitz F. The Pseudomonas phytotoxin coronatine mimics octadecanoid signalling molecules of higher plants. FEBS Lett. 1994 May 23;345(1):9–13. [PubMed]
  • Winans SC, Ebert PR, Stachel SE, Gordon MP, Nester EW. A gene essential for Agrobacterium virulence is homologous to a family of positive regulatory loci. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8278–8282. [PMC free article] [PubMed]
  • Xiao Y, Heu S, Yi J, Lu Y, Hutcheson SW. Identification of a putative alternate sigma factor and characterization of a multicomponent regulatory cascade controlling the expression of Pseudomonas syringae pv. syringae Pss61 hrp and hrmA genes. J Bacteriol. 1994 Feb;176(4):1025–1036. [PMC free article] [PubMed]
  • Xiao Y, Lu Y, Heu S, Hutcheson SW. Organization and environmental regulation of the Pseudomonas syringae pv. syringae 61 hrp cluster. J Bacteriol. 1992 Mar;174(6):1734–1741. [PMC free article] [PubMed]
  • Yoshikawa H, Kazami J, Yamashita S, Chibazakura T, Sone H, Kawamura F, Oda M, Isaka M, Kobayashi Y, Saito H. Revised assignment for the Bacillus subtilis spo0F gene and its homology with spo0A and with two Escherichia coli genes. Nucleic Acids Res. 1986 Jan 24;14(2):1063–1072. [PMC free article] [PubMed]
  • Young SA, Park SK, Rodgers C, Mitchell RE, Bender CL. Physical and functional characterization of the gene cluster encoding the polyketide phytotoxin coronatine in Pseudomonas syringae pv. glycinea. J Bacteriol. 1992 Mar;174(6):1837–1843. [PMC free article] [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...