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Structural basis of pathogen recognition by an integrated HMA domain in a plant NLR immune receptor.

Maqbool A, Saitoh H, Franceschetti M, Stevenson CE, Uemura A, Kanzaki H, Kamoun S, Terauchi R, Banfield MJ - Elife (2015)

Bottom Line: Plants have evolved intracellular immune receptors to detect pathogen proteins known as effectors.How these immune receptors detect effectors remains poorly understood.The crystal structure of the Pikp-HMA/AVR-PikD complex enabled design of mutations to alter protein interaction in yeast and in vitro, and perturb effector-mediated response both in a rice cultivar containing Pikp and upon expression of AVR-PikD and Pikp in the model plant Nicotiana benthamiana.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Chemistry, John Innes Centre, Norwich, United Kingdom.

ABSTRACT
Plants have evolved intracellular immune receptors to detect pathogen proteins known as effectors. How these immune receptors detect effectors remains poorly understood. Here we describe the structural basis for direct recognition of AVR-Pik, an effector from the rice blast pathogen, by the rice intracellular NLR immune receptor Pik. AVR-PikD binds a dimer of the Pikp-1 HMA integrated domain with nanomolar affinity. The crystal structure of the Pikp-HMA/AVR-PikD complex enabled design of mutations to alter protein interaction in yeast and in vitro, and perturb effector-mediated response both in a rice cultivar containing Pikp and upon expression of AVR-PikD and Pikp in the model plant Nicotiana benthamiana. These data reveal the molecular details of a recognition event, mediated by a novel integrated domain in an NLR, which initiates a plant immune response and resistance to rice blast disease. Such studies underpin novel opportunities for engineering disease resistance to plant pathogens in staple food crops.

No MeSH data available.


SDS-PAGE of AVR-PikD mutant proteins and SPR sensorgrams.(A) AVR-PikD mutants prepared for SPR analysis. (B) Single-cycle kinetics data for the interaction of Pikp-HMA (analyte) with AVR-PikD mutants. This data was used to generate the binding curves shown in Figure 4B.DOI:http://dx.doi.org/10.7554/eLife.08709.019
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fig4s2: SDS-PAGE of AVR-PikD mutant proteins and SPR sensorgrams.(A) AVR-PikD mutants prepared for SPR analysis. (B) Single-cycle kinetics data for the interaction of Pikp-HMA (analyte) with AVR-PikD mutants. This data was used to generate the binding curves shown in Figure 4B.DOI:http://dx.doi.org/10.7554/eLife.08709.019

Mentions: (A) Y2H assays showing the binding of AVR-PikD mutants to Pikp-HMA using two read-outs, growth on–Leu-Trp-His+3AT (-LTH) plates and the X-gal assay. (B) Binding curves derived from Surface Plasmon Resonance single-cycle kinetics data for Pikp-HMA binding to AVR-PikD and AVR-PikD mutants, Kd values are shown where determined (ND = Not Determined, NB = No Binding). The sensorgrams of the data used to derive these curves are shown in Figure 4—figure supplement 2B.


Structural basis of pathogen recognition by an integrated HMA domain in a plant NLR immune receptor.

Maqbool A, Saitoh H, Franceschetti M, Stevenson CE, Uemura A, Kanzaki H, Kamoun S, Terauchi R, Banfield MJ - Elife (2015)

SDS-PAGE of AVR-PikD mutant proteins and SPR sensorgrams.(A) AVR-PikD mutants prepared for SPR analysis. (B) Single-cycle kinetics data for the interaction of Pikp-HMA (analyte) with AVR-PikD mutants. This data was used to generate the binding curves shown in Figure 4B.DOI:http://dx.doi.org/10.7554/eLife.08709.019
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4547098&req=5

fig4s2: SDS-PAGE of AVR-PikD mutant proteins and SPR sensorgrams.(A) AVR-PikD mutants prepared for SPR analysis. (B) Single-cycle kinetics data for the interaction of Pikp-HMA (analyte) with AVR-PikD mutants. This data was used to generate the binding curves shown in Figure 4B.DOI:http://dx.doi.org/10.7554/eLife.08709.019
Mentions: (A) Y2H assays showing the binding of AVR-PikD mutants to Pikp-HMA using two read-outs, growth on–Leu-Trp-His+3AT (-LTH) plates and the X-gal assay. (B) Binding curves derived from Surface Plasmon Resonance single-cycle kinetics data for Pikp-HMA binding to AVR-PikD and AVR-PikD mutants, Kd values are shown where determined (ND = Not Determined, NB = No Binding). The sensorgrams of the data used to derive these curves are shown in Figure 4—figure supplement 2B.

Bottom Line: Plants have evolved intracellular immune receptors to detect pathogen proteins known as effectors.How these immune receptors detect effectors remains poorly understood.The crystal structure of the Pikp-HMA/AVR-PikD complex enabled design of mutations to alter protein interaction in yeast and in vitro, and perturb effector-mediated response both in a rice cultivar containing Pikp and upon expression of AVR-PikD and Pikp in the model plant Nicotiana benthamiana.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Chemistry, John Innes Centre, Norwich, United Kingdom.

ABSTRACT
Plants have evolved intracellular immune receptors to detect pathogen proteins known as effectors. How these immune receptors detect effectors remains poorly understood. Here we describe the structural basis for direct recognition of AVR-Pik, an effector from the rice blast pathogen, by the rice intracellular NLR immune receptor Pik. AVR-PikD binds a dimer of the Pikp-1 HMA integrated domain with nanomolar affinity. The crystal structure of the Pikp-HMA/AVR-PikD complex enabled design of mutations to alter protein interaction in yeast and in vitro, and perturb effector-mediated response both in a rice cultivar containing Pikp and upon expression of AVR-PikD and Pikp in the model plant Nicotiana benthamiana. These data reveal the molecular details of a recognition event, mediated by a novel integrated domain in an NLR, which initiates a plant immune response and resistance to rice blast disease. Such studies underpin novel opportunities for engineering disease resistance to plant pathogens in staple food crops.

No MeSH data available.