Functional studies of the bacterial avirulence protein AvrPto by mutational analysis

Molecular Plant-microbe Interactions : MPMI
J H ChangRichard W Michelmore

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 ...Continue Reading

References

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Citations

Feb 8, 2002·Trends in Plant Science·Renier A L Van der HoornMatthieu H A J Joosten
Jul 1, 2002·Molecular Plant Pathology·Adam J Bogdanove
Sep 25, 2003·Annual Review of Plant Biology·Gregory B MartinGuido Sessa
Oct 7, 2003·Annual Review of Phytopathology·Kerry F Pedley, Gregory B Martin
Sep 4, 2007·The Plant Journal : for Cell and Molecular Biology·Fangming XiaoGregory B Martin
Jul 10, 2004·Structure·David A Horita
Apr 5, 2014·Chemical Reviews·Macarena Marín, Thomas Ott
Mar 26, 2005·The Journal of Biological Chemistry·Adriana J BernalRichard W Michelmore

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