Functional studies of the bacterial avirulence protein AvrPto by mutational analysis

J. H. Chang, C. M. Tobias, B. J. Staskawicz, Richard W Michelmore

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Pseudomonas syringae pathovars expressing avrPto are avirulent on plants expressing the resistance gene Pto. Over 85 mutants of avrPto were generated with multiple strategies, and several assays were used to characterize AvrPto function. Only a core of 95 amino acids of the 164 residues was sufficient for binding Pto in the yeast two-hybrid system. Only nine of 65 mutant proteins of AvrPto with amino acid substitutions, created in planta and in vitro, did not interact with Pto in the Gal4 yeast two-hybrid system, suggesting that AvrPto can tolerate many nonconservative substitutions and still interact with Pto. These nine and 12 additional substitution mutants of AvrPto were characterized further. The ability to elicit a hypersensitive response and the effect on pathogenesis in planta for these 21 mutants of AvrPto were strongly correlated with recognition by Pto in the yeast two-hybrid system. Analyses of two proteins with substitutions H54P or D52G/L65P indicated that these residues may be required for delivery into the host cell and protein stability in the bacterial cytoplasm, respectively. The mutants that no longer interacted with Pto and had modified activities in planta were predicted to have changes in their secondary structure.

Original languageEnglish (US)
Pages (from-to)451-459
Number of pages9
JournalMolecular Plant-Microbe Interactions
Volume14
Issue number4
StatePublished - 2001

Fingerprint

Two-Hybrid System Techniques
bacterial proteins
Bacterial Proteins
Substitution reactions
two hybrid system techniques
Hybrid systems
Yeast
mutants
Plantae
Pseudomonas syringae
yeasts
Protein Stability
Mutant Proteins
Amino Acid Substitution
Amino Acids
Cytoplasm
proteins
amino acid substitution
hypersensitive response
Assays

ASJC Scopus subject areas

  • Microbiology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology

Cite this

Chang, J. H., Tobias, C. M., Staskawicz, B. J., & Michelmore, R. W. (2001). Functional studies of the bacterial avirulence protein AvrPto by mutational analysis. Molecular Plant-Microbe Interactions, 14(4), 451-459.

Functional studies of the bacterial avirulence protein AvrPto by mutational analysis. / Chang, J. H.; Tobias, C. M.; Staskawicz, B. J.; Michelmore, Richard W.

In: Molecular Plant-Microbe Interactions, Vol. 14, No. 4, 2001, p. 451-459.

Research output: Contribution to journalArticle

Chang, JH, Tobias, CM, Staskawicz, BJ & Michelmore, RW 2001, 'Functional studies of the bacterial avirulence protein AvrPto by mutational analysis', Molecular Plant-Microbe Interactions, vol. 14, no. 4, pp. 451-459.
Chang, J. H. ; Tobias, C. M. ; Staskawicz, B. J. ; Michelmore, Richard W. / Functional studies of the bacterial avirulence protein AvrPto by mutational analysis. In: Molecular Plant-Microbe Interactions. 2001 ; Vol. 14, No. 4. pp. 451-459.
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