Home LiteratureArticle Details
PMID: 10430949 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The Xanthomonas Hrp type III system secretes proteins from plant and mammalian bacterial pathogens.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 16 ·1999-08-03 ·Pages 9368-73

Rossier O, Wengelnik K, Hahn K, Bonas U

Abstract

Studies of essential pathogenicity determinants in Gram-negative bacteria have revealed the conservation of type III protein secretion systems that allow delivery of virulence factors into host cells from plant and animal pathogens. Ten of 21 Hrp proteins of the plant pathogen Xanthomonas campestris pv. vesicatoria have been suggested to be part of a type III machinery. Here, we report the hrp-dependent secretion of two avirulence proteins, AvrBs3 and AvrRxv, by X. campestris pv. vesicatoria strains that constitutively express hrp genes. Secretion occurred without leakage of a cytoplasmic marker in minimal medium containing BSA, at pH 5.4. Secretion was strictly hrp-dependent because a mutant carrying a deletion in hrcV, a conserved hrp gene, did not secrete AvrBs3 and AvrRxv. Moreover, the Hrp system of X. campestris pv. vesicatoria was able to secrete proteins from two other plant pathogens: PopA, a protein secreted via the Hrp system in Ralstonia solanacearum, and AvrB, an avirulence protein from Pseudomonas syringae pv. glycinea. Interestingly, X. campestris pv. vesicatoria also secreted YopE, a type III-secreted cytotoxin of the mammalian pathogen Yersinia pseudotuberculosis in a hrp-dependent manner. YerA, a YopE-specific chaperone, was required for YopE stability but not for secretion in X. campestris pv. vesicatoria. Our results demonstrate the functional conservation of the type III system of X. campestris for secretion of proteins from both plant and mammalian pathogens and imply recognition of their respective secretion signals.

MeSH Terms
Animals Bacterial Outer Membrane Proteins/genetics Bacterial Proteins/biosynthesis,genetics Bacterial Toxins/genetics Cloning, Molecular Gene Expression Regulation, Bacterial Glucuronidase/biosynthesis,genetics Mammals Mutagenesis Plants/microbiology Pseudomonas/genetics Recombinant Fusion Proteins/biosynthesis Sequence Deletion Transcription Activator-Like Effectors Virulence Xanthomonas campestris/genetics,pathogenicity,physiology Yersinia pseudotuberculosis/genetics
Chemicals
Bacterial Outer Membrane Proteins Bacterial Proteins Bacterial Toxins Recombinant Fusion Proteins Transcription Activator-Like Effectors avrRxv protein, Xanthomonas campestris yopE protein, Yersinia avrBs3-2 protein, Xanthomonas campestris Glucuronidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rossier O
Centre National de la Recherche Scientifique, Institut des Sciences Végétales, Avenue de la Terrasse, 91190 Gif-sur-Yvette Cedex, France.
Wengelnik K
Hahn K
Bonas U
References (48)
48 references, click to expand
  1. Genetic and structural characterization of the avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria.
    Mol Gen Genet. 1989 Jul;218(1):127-36 PMID: 2550761
  2. Avirulence gene avrRxv from Xanthomonas campestris pv. vesicatoria specifies resistance on tomato line Hawaii 7998.
    Mol Plant Microbe Interact. 1993 Sep-Oct;6(5):616-27 PMID: 8274773
  3. A complementation analysis of the restriction and modification of DNA in Escherichia coli.
    J Mol Biol. 1969 May 14;41(3):459-72 PMID: 4896022
  4. Intracellular targeting of the Yersinia YopE cytotoxin in mammalian cells induces actin microfilament disruption.
    Infect Immun. 1991 Dec;59(12):4562-9 PMID: 1937815
  5. PopA1, a protein which induces a hypersensitivity-like response on specific Petunia genotypes, is secreted via the Hrp pathway of Pseudomonas solanacearum.
    EMBO J. 1994 Feb 1;13(3):543-53 PMID: 8313899
  6. A mRNA signal for the type III secretion of Yop proteins by Yersinia enterocolitica.
    Science. 1997 Nov 7;278(5340):1140-3 PMID: 9353199
  7. The A. thaliana disease resistance gene RPS2 encodes a protein containing a nucleotide-binding site and leucine-rich repeats.
    Cell. 1994 Sep 23;78(6):1089-99 PMID: 7923358
  8. A gene from Xanthomonas campestris pv. vesicatoria that determines avirulence in tomato is related to avrBs3.
    Mol Plant Microbe Interact. 1991 Nov-Dec;4(6):628-32 PMID: 1804405
  9. Expression of the avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria is not under the control of hrp genes and is independent of plant factors.
    J Bacteriol. 1991 Nov;173(22):7142-50 PMID: 1938914
  10. DspA, an essential pathogenicity factor of Erwinia amylovora showing homology with AvrE of Pseudomonas syringae, is secreted via the Hrp secretion pathway in a DspB-dependent way.
    Mol Microbiol. 1997 Dec;26(5):1057-69 PMID: 9426142
  11. Bacterial blight of soybean: regulation of a pathogen gene determining host cultivar specificity.
    Science. 1989 Sep 22;245(4924):1374-7 PMID: 2781284
  12. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.
    EMBO J. 1987 Dec 20;6(13):3901-7 PMID: 3327686
  13. The Salmonella typhimurium invasion genes invF and invG encode homologues of the AraC and PulD family of proteins.
    Mol Microbiol. 1994 Aug;13(4):555-68 PMID: 7997169
  14. HrpG, a key hrp regulatory protein of Xanthomonas campestris pv. vesicatoria is homologous to two-component response regulators.
    Mol Plant Microbe Interact. 1996 Nov;9(8):704-12 PMID: 8870269
  15. Production of single-stranded plasmid DNA.
    Methods Enzymol. 1987;153:3-11 PMID: 3323803
  16. The Yersinia deadly kiss.
    J Bacteriol. 1998 Nov;180(21):5495-504 PMID: 9791096
  17. Characterization of the Pseudomonas syringae pv. tomato AvrRpt2 protein: demonstration of secretion and processing during bacterial pathogenesis.
    Mol Microbiol. 1999 Jun;32(5):927-41 PMID: 10361296
  18. Functional conservation of the secretion and translocation machinery for virulence proteins of yersiniae, salmonellae and shigellae.
    EMBO J. 1995 Sep 1;14(17):4187-95 PMID: 7556059
  19. HrpXv, an AraC-type regulator, activates expression of five of the six loci in the hrp cluster of Xanthomonas campestris pv. vesicatoria.
    J Bacteriol. 1996 Jun;178(12):3462-9 PMID: 8655542
  20. Recognition of the bacterial avirulence protein AvrBs3 occurs inside the host plant cell.
    Cell. 1996 Dec 27;87(7):1307-16 PMID: 8980236
  21. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  22. Sequence and expression analysis of the hrpB pathogenicity operon of Xanthomonas campestris pv. vesicatoria which encodes eight proteins with similarity to components of the Hrp, Ysc, Spa, and Fli secretion systems.
    Mol Plant Microbe Interact. 1995 Nov-Dec;8(6):845-54 PMID: 8664494
  23. Determinants of pathogenicity in Xanthomonas campestris pv. vesicatoria are related to proteins involved in secretion in bacterial pathogens of animals.
    Mol Plant Microbe Interact. 1992 Sep-Oct;5(5):390-6 PMID: 1472717
  24. hpaA mutants of Xanthomonas campestris pv. vesicatoria are affected in pathogenicity but retain the ability to induce host-specific hypersensitive reaction.
    Mol Microbiol. 1998 Sep;29(6):1379-90 PMID: 9781876
  25. The Pseudomonas syringae pv. tomato HrpW protein has domains similar to harpins and pectate lyases and can elicit the plant hypersensitive response and bind to pectate.
    J Bacteriol. 1998 Oct;180(19):5211-7 PMID: 9748456
  26. Yersinia enterocolitica induces apoptosis in macrophages by a process requiring functional type III secretion and translocation mechanisms and involving YopP, presumably acting as an effector protein.
    Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12638-43 PMID: 9356502
  27. Genetic analysis of the yopE region of Yersinia spp.: identification of a novel conserved locus, yerA, regulating yopE expression.
    J Bacteriol. 1990 Mar;172(3):1547-55 PMID: 2307658
  28. Erwinia amylovora secretes harpin via a type III pathway and contains a homolog of yopN of Yersinia spp.
    J Bacteriol. 1996 Mar;178(6):1720-30 PMID: 8626302
  29. Unified nomenclature for broadly conserved hrp genes of phytopathogenic bacteria.
    Mol Microbiol. 1996 May;20(3):681-3 PMID: 8736546
  30. Characterization of a gene from a tomato pathogen determining hypersensitive resistance in non-host species and genetic analysis of this resistance in bean.
    Proc Natl Acad Sci U S A. 1988 Sep;85(18):6743-7 PMID: 16593982
  31. HrpW of Erwinia amylovora, a new harpin that contains a domain homologous to pectate lyases of a distinct class.
    J Bacteriol. 1998 Oct;180(19):5203-10 PMID: 9748455
  32. A secreted Salmonella protein with homology to an avirulence determinant of plant pathogenic bacteria.
    Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9887-92 PMID: 9275221
  33. The chaperone-like protein YerA of Yersinia pseudotuberculosis stabilizes YopE in the cytoplasm but is dispensible for targeting to the secretion loci.
    Mol Microbiol. 1995 May;16(4):635-47 PMID: 7476159
  34. Molecular characterization of cloned avirulence genes from race 0 and race 1 of Pseudomonas syringae pv. glycinea.
    J Bacteriol. 1987 Dec;169(12):5789-94 PMID: 2824447
  35. Nonpolar mutagenesis of the ipa genes defines IpaB, IpaC, and IpaD as effectors of Shigella flexneri entry into epithelial cells.
    J Bacteriol. 1993 Sep;175(18):5899-906 PMID: 8376337
  36. Type III protein secretion systems in bacterial pathogens of animals and plants.
    Microbiol Mol Biol Rev. 1998 Jun;62(2):379-433 PMID: 9618447
  37. Expression of Erwinia amylovora hrp genes in response to environmental stimuli.
    J Bacteriol. 1992 Mar;174(6):1875-82 PMID: 1372313
  38. Hrp pilus: an hrp-dependent bacterial surface appendage produced by Pseudomonas syringae pv. tomato DC3000.
    Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3459-64 PMID: 9096416
  39. Characterization of the operon encoding the YpkA Ser/Thr protein kinase and the YopJ protein of Yersinia pseudotuberculosis.
    J Bacteriol. 1994 Aug;176(15):4543-8 PMID: 8045884
  40. In vitro insertional mutagenesis with a selectable DNA fragment.
    Gene. 1984 Sep;29(3):303-13 PMID: 6237955
  41. Expression and localization of HrpA1, a protein of Xanthomonas campestris pv. vesicatoria essential for pathogenicity and induction ofthe hypersensitive reaction.
    J Bacteriol. 1996 Feb;178(4):1061-9 PMID: 8576039
  42. A cloned Erwinia chrysanthemi Hrp (type III protein secretion) system functions in Escherichia coli to deliver Pseudomonas syringae Avr signals to plant cells and to secrete Avr proteins in culture.
    Proc Natl Acad Sci U S A. 1998 Aug 18;95(17):10206-11 PMID: 9707625
  43. The type III (Hrp) secretion pathway of plant pathogenic bacteria: trafficking harpins, Avr proteins, and death.
    J Bacteriol. 1997 Sep;179(18):5655-62 PMID: 9294418
  44. Pseudomonas syringae pv. syringae harpinPss: a protein that is secreted via the Hrp pathway and elicits the hypersensitive response in plants.
    Cell. 1993 Jul 2;73(7):1255-66 PMID: 8324821
  45. Expression of the Pseudomonas syringae avirulence protein AvrB in plant cells alleviates its dependence on the hypersensitive response and pathogenicity (Hrp) secretion system in eliciting genotype-specific hypersensitive cell death.
    Plant Cell. 1996 Jul;8(7):1095-105 PMID: 8768370
  46. The avrRpm1 gene of Pseudomonas syringae pv. maculicola is required for virulence on Arabidopsis.
    Mol Plant Microbe Interact. 1995 May-Jun;8(3):444-53 PMID: 7655064
  47. Cloning of genes involved in pathogenicity of Xanthomonas campestris pv. campestris using the broad host range cosmid pLAFR1.
    EMBO J. 1984 Dec 20;3(13):3323-8 PMID: 16453595
  48. A Xanthomonas Pathogenicity Locus Is Induced by Sucrose and Sulfur-Containing Amino Acids.
    Plant Cell. 1992 Jan;4(1):79-86 PMID: 12297631
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.16.9368
Published
1999-08-03
Pages
9368-73
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC17789
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.16.9368 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