Structural elucidation and functional characterization of the Hyaloperonospora arabidopsidis effector protein ATR13

Lauriebeth Leonelli, Jeffery Pelton, Allyn Schoeffler, Douglas Dahlbeck, James M Berger, David E. Wemmer, Brian Staskawicz

Research output: Contribution to journalArticle

Abstract

The oomycete Hyaloperonospora arabidopsidis (Hpa) is the causal agent of downy mildew on the model plant Arabidopsis thaliana and has been adapted as a model system to investigate pathogen virulence strategies and plant disease resistance mechanisms. Recognition of Hpa infection occurs when plant resistance proteins (R-genes) detect the presence or activity of pathogen-derived protein effectors delivered to the plant host. This study examines the Hpa effector ATR13 Emco5 and its recognition by RPP13-Nd, the cognate R-gene that triggers programmed cell death (HR) in the presence of recognized ATR13 variants. Herein, we use NMR to solve the backbone structure of ATR13 Emco5, revealing both a helical domain and a disordered internal loop. Additionally, we use site-directed and random mutagenesis to identify several amino acid residues involved in the recognition response conferred by RPP13-Nd. Using our structure as a scaffold, we map these residues to one of two surface-exposed patches of residues under diversifying selection. Exploring possible roles of the disordered region within the ATR13 structure, we perform domain swapping experiments and identify a peptide sequence involved in nucleolar localization. We conclude that ATR13 is a highly dynamic protein with no clear structural homologues that contains two surface-exposed patches of polymorphism, only one of which is involved in RPP13-Nd recognition specificity.

Original languageEnglish (US)
Article numbere1002428
JournalPLoS Pathogens
Volume7
Issue number12
DOIs
StatePublished - Dec 2011
Externally publishedYes

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vpr Genes
Oomycetes
Plant Diseases
Plant Proteins
Disease Resistance
Site-Directed Mutagenesis
Arabidopsis
Virulence
Proteins
Cell Death
Amino Acids
Peptides
Infection

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Structural elucidation and functional characterization of the Hyaloperonospora arabidopsidis effector protein ATR13. / Leonelli, Lauriebeth; Pelton, Jeffery; Schoeffler, Allyn; Dahlbeck, Douglas; Berger, James M; Wemmer, David E.; Staskawicz, Brian.

In: PLoS Pathogens, Vol. 7, No. 12, e1002428, 12.2011.

Research output: Contribution to journalArticle

Leonelli, Lauriebeth ; Pelton, Jeffery ; Schoeffler, Allyn ; Dahlbeck, Douglas ; Berger, James M ; Wemmer, David E. ; Staskawicz, Brian. / Structural elucidation and functional characterization of the Hyaloperonospora arabidopsidis effector protein ATR13. In: PLoS Pathogens. 2011 ; Vol. 7, No. 12.
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