Skip to main content
SpringerLink
Log in

Structural characterization of protein secretion genes of the bacterial phytopathogen Xanthomonas campestris pathovar campestris: relatedness to secretion systems of other gram-negative bacteria

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The nucleotide sequence was determined of a 5.3 kb region of the Xanthomonas campestris pathovar campestris genome carrying a gene cluster encoding protein secretion and pathogenicity functions. A putative promoter sequence and five open reading frames (ORF) which may be part of an operon were revealed. The five predicted primary translation products comprise 531, 390, 147, 169 and 138 amino acids with Mr values of 58854, 42299, 15548, 18214 and 15108 respectively. A sixth, partial ORF is also present. Between ORF1 and ORF2 is a sequence of unknown function showing 7 by duplications. The deduced amino acid sequence of ORF1 is related to the Klebsiella pneumoniae PulE protein, to the Bacillus subtilis ComG ORF1 and to the Agrobacterium tumefaciens VirB ORF11 products. In addition, the deduced amino acid sequence of ORF2 showed homology to the Pu1F and to the ComG ORF2 products. The proteins encoded by ORF3, 4 and 5 showed amino acid homology to PulG, H and I products respectively. The proteins encoded by ORF2, 3, 4 and 5 showed significant hydrophobic domains which may represent membrane-spanning regions. By contrast the protein encoded by ORF1 was largely hydrophilic and had two putative nucleoside triphosphate binding sites.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Albano M, Breitling R, Dubnau DA (1989) Nucleotide sequence and genetic organization of the Bacillus subtilis comG operon. J Bacteriol 171:5386–5404

    Google Scholar 

  • Bibb MJ, Findlay RP, Johnson MW (1984) The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of proteincoding sequences. Gene 30:157–166

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Boccara M, Expert D, Diolez A, Bertheau Y, Kotoujansky A (1986) Erwinia chrysanthemi as a model to study pathogenicity and as a biotechnological tool. 3rd International Symposium Molecular Genetics of Plant-Microbe Interactions, Montreal, Abstract p 76

  • Daniels MJ, Barber CE, Turner PC, Cleary WG, Sawczyc MK (1984a) Isolation of mutants of Xanthomonas campestris pv. campestris showing altered pathogenicity. J Gen Microbiol 130:2447–2455

    Google Scholar 

  • Daniels MJ, Barber CE, Turner PC, Sawczyc MK, Byrde RJW, Fielding AH (1984b) Cloning of genes involved in pathogenicity of Xanthomonas campestris pv. campestris using the broad host range cosmid pLAFR1. EMBO J 3:3323–3328

    Google Scholar 

  • D'Enfert C, Ryter A, Pusgley AP (1987) Cloning and expression in Escherichia coli of the Klebsiella pneumoniae genes for production, surface localisation and secretion of the lipoprotein pullulanase. EMBO J 6:3531–3538

    Google Scholar 

  • Dow JM, Scofield G, Trafford K, Turner PC, Daniels MJ (1987) A gene cluster in Xanthomonas campestris pv. campestris required for pathogenicity controls the excretion of polygalacturonate lyase and other enzymes. Physiol Mol Plant Pathol 31:261–271

    Google Scholar 

  • Dow JM, Daniels MJ, Dums F, Turner PC, Gough C (1989) Genetic and biochemical analysis of protein export from Xanthomonas campestris. J Cell Sci (Suppl) 11:59–72

    Google Scholar 

  • Filloux A, Bally M, Ball G, Akrim M, Tommassen J, Lazdunski A (1990) Protein secretion in Gram-negative bacteria: transport across the outer membrane involves common mechanisms in different bacteria. EMBO J 9:4323–4329

    Google Scholar 

  • Fürste JP, Pansegrau W, Frank R, Blacker H, Scholz P, Bagdasarian M, Lanka E (1986) Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene 48:119–131

    Google Scholar 

  • Gough CL, Dow JM, Keen J, Henrissat B, Daniels MJ (1990) Nucleotide sequence of the engX gene encoding the major endoglucanase of Xanthomonas campestris pathovar campestris. Gene 89:53–59

    Google Scholar 

  • Gray L, Baker K, Kenny B, Mackman N, Haigh R, Holland IB (1989) A novel C-terminal signal sequence targets Escherichia coli haemolysin directly to the medium. J Cell Sci (Suppl) 11:45–57

    Google Scholar 

  • He SY, Lindeberg M, Chatterjee AK, Collmer A (1991) Cloned Erwinia chrysanthemi out genes enable Escherichia coli to selectively secrete a diverse family of heterologous proteins to its milieu. Proc Natl Acad Sci USA 88:1079–1083

    Google Scholar 

  • Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    Google Scholar 

  • Ji J, Hugovieux-Cotte-Pattat N, Robert-Baudouy J (1989) Molecular cloning of the outJ gene involved in pectate lyase secretion by Erwinia chrysanthemi. Mol Microbiol 3:285–293

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Laminck AA, Zielgelhaffer T, Georgopoulos C, Plückthun A (1990) The Escherichia coli heat shock proteins GroEL and GroES modulate the folding of the β-lactamase precursor. EMBO J 9:2315–2319

    Google Scholar 

  • Letoffe S, Delepelaire P, Wandersman C (1990) Protease secretion by Erwinia chrysanthemi: the specific secretion functions are analogous to those of Escherichia coli α-haemolysin. EMBO 79:1375–1382

    Google Scholar 

  • Liu Y-N, Tang J-L, Clarke BR, Dow JM, Daniels MJ (1990) A multi-purpose broad host range cloning vector and its use to characterize an extracellular protease gene of Xanthomonas campestris pathovar campestris. Mol Gen Genet 220:433–440

    Google Scholar 

  • Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double digest restriction fragments. Gene 19:269–276

    Google Scholar 

  • Model P, Russel M (1990) Prokaryotic secretion. Cell 61:739–741

    Google Scholar 

  • Murata H, Fons M, Chatterjee A, Collmer A, Chatterjee AK (1990) Characterization of transposon insertion Out− mutants of Erwinia carotovora subsp. carotovora defective in enzyme export and of a DNA segment that complements out mutations in E. carotovora subsp. carotovora, E. carotovora subsp. atroseptica, and Erwinia chrysanthemi. J Bacteriol 172:2970–2978

    Google Scholar 

  • Pugsley AP, Reyss I (1990) Five genes at the 3′ end of the Klebsiella pneumoniae puIC operon are required for pullulanase secretion. Mol Microbiol 4:365–379

    Google Scholar 

  • Pugsley AP, d'Enfert C, Reyss I, Kornacker MG (1990) Genetics of extracellular protein secretion by Gram-negative bacteria. Annu Rev Genet 24:67–90

    Google Scholar 

  • Reyss I, Pugsley AP (1990) Five additional genes in the pulC-O operon of the Gram-negative bacterium Klebsiella oxytoca UNF5023 which are required for pullulanase secretion. Mol Gen Genet 222:176–184

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA: Complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Thompson DV, Melchers LS, Idler KB, Schilperoort RA, Hooykaas PJJ (1988) Analysis of the complete nucleotide sequence of the Agrobacterium tumefaciens virB operon. Nucleic Acids Res 16:4621–4636

    Google Scholar 

  • Turner P, Barber C, Daniels M (1984) Behaviour of the transposons Tn5 and Tn7 in Xanthomonas campestris pv. campestris. Mol Gen Genet 195:101–107

    Google Scholar 

  • Turner P, Barber C, Daniels M (1985) Evidence for clustered pathogenicity genes in Xanthomonas campestris pv. campestris. Mol Gen Genet 199:338–343

    Google Scholar 

  • Walker JE, Saraste M, Runswick MJ, Gay NJ (1982) Distantly related sequences in the α- and β-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951

    Google Scholar 

  • Ward JE, Akiyoshi DE, Regier D, Datta A, Gordon MP, Nester EW (1988) Characterization of the virB operon from an Agrobacterium tumefaciens Ti plasmid. J Biol Chem 263:5804–5814

    Google Scholar 

  • Williams PH (1980) Black rot: a continuing threat to world crucifers. Plant Dis 64:736–74

    Google Scholar 

Download references

Author information

Author notes

  1. Franz Dums

    Present address: Lehrstuhl für Genetik, Universität Bayreuth, Universitätsstr. 30, D-8580, Bayreuth, Germany

  2. J. Maxwell Dow & Michael J. Daniels

    Present address: The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK

Authors and Affiliations

  1. John Innes Institute, Colney Lane, Norwich, NR4 7UH, UK

    Franz Dums, J. Maxwell Dow & Michael J. Daniels

Authors
  1. Franz Dums
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Maxwell Dow
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael J. Daniels
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by W. Goebel

The nucleotide sequence data in this paper have been deposited in the EMBL, Genbank and DDBJ nucleotide sequence databases under the accession number X59079

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dums, F., Dow, J.M. & Daniels, M.J. Structural characterization of protein secretion genes of the bacterial phytopathogen Xanthomonas campestris pathovar campestris: relatedness to secretion systems of other gram-negative bacteria. Molec. Gen. Genet. 229, 357–364 (1991). https://doi.org/10.1007/BF00267456

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00267456

Key words

Search

Navigation