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Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato.

Publication ,  Journal Article
Lee, YH; Kolade, OO; Nomura, K; Arvidson, DN; He, SY
Published in: The Journal of biological chemistry
June 2005

The Hrp pilus plays an essential role in the long-distance type III translocation of effector proteins from bacteria into plant cells. HrpA is the structural subunit of the Hrp pilus in Pseudomonas syringae pv. tomato (Pst) DC3000. Little is known about the molecular features in the HrpA protein for pilus assembly or for transporting effector proteins. From previous collections of nonfunctional HrpA derivatives that carry random pentapeptide insertions or single amino acid mutations, we identified several dominant-negative mutants that blocked the ability of wild-type Pst DC3000 to elicit host responses. The dominant-negative phenotype was correlated with the disappearance of the Hrp pilus in culture and inhibition of wild-type HrpA protein self-assembly in vitro. Dominant-negative HrpA mutants can be grouped into two functional classes: one class exerted a strong dominant-negative effect on the secretion of effector proteins AvrPto and HopPtoM in culture, and the other did not. The two classes of mutant HrpA proteins carry pentapeptide insertions in discrete regions, which are interrupted by insertions without a dominant-negative effect. These results enable prediction of possible subunit-subunit interaction sites in the assembly of the Hrp pilus and suggest the usefulness of dominant-negative mutants in dissection of the role of the wild-type HrpA protein in various stages of type III translocation: protein exit across the bacterial cell wall, the assembly and/or stabilization of the Hrp pilus in the extracellular space, and Hrp pilus-mediated long-distance transport beyond the bacterial cell wall.

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Published In

The Journal of biological chemistry

DOI

EISSN

1083-351X

ISSN

0021-9258

Publication Date

June 2005

Volume

280

Issue

22

Start / End Page

21409 / 21417

Related Subject Headings

  • RNA Helicases
  • Pseudomonas syringae
  • Protein Transport
  • Protein Structure, Tertiary
  • Protein Binding
  • Phenotype
  • Peptides
  • Mutation
  • Molecular Sequence Data
  • Microscopy, Electron, Transmission
 

Citation

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Lee, Y. H., Kolade, O. O., Nomura, K., Arvidson, D. N., & He, S. Y. (2005). Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato. The Journal of Biological Chemistry, 280(22), 21409–21417. https://doi.org/10.1074/jbc.m500972200
Lee, Yong Hoon, Olatomirin O. Kolade, Kinya Nomura, Dennis N. Arvidson, and Sheng Yang He. “Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato.The Journal of Biological Chemistry 280, no. 22 (June 2005): 21409–17. https://doi.org/10.1074/jbc.m500972200.
Lee YH, Kolade OO, Nomura K, Arvidson DN, He SY. Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato. The Journal of biological chemistry. 2005 Jun;280(22):21409–17.
Lee, Yong Hoon, et al. “Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato.The Journal of Biological Chemistry, vol. 280, no. 22, June 2005, pp. 21409–17. Epmc, doi:10.1074/jbc.m500972200.
Lee YH, Kolade OO, Nomura K, Arvidson DN, He SY. Use of dominant-negative HrpA mutants to dissect Hrp pilus assembly and type III secretion in Pseudomonas syringae pv. tomato. The Journal of biological chemistry. 2005 Jun;280(22):21409–21417.

Published In

The Journal of biological chemistry

DOI

EISSN

1083-351X

ISSN

0021-9258

Publication Date

June 2005

Volume

280

Issue

22

Start / End Page

21409 / 21417

Related Subject Headings

  • RNA Helicases
  • Pseudomonas syringae
  • Protein Transport
  • Protein Structure, Tertiary
  • Protein Binding
  • Phenotype
  • Peptides
  • Mutation
  • Molecular Sequence Data
  • Microscopy, Electron, Transmission