A translocation signal for delivery of oomycete effector proteins inside host plant cells

Stephen C. Whisson, Petra C. Boevink, Lucy Moleleki, Anna O. Avrova, Juan G. Morales, Eleanor M. Gilroy, Miles R. Armstrong, Severine Grouffaud, Pieter van West, Sean Chapman, Ingo Hein, Ian K. Toth, Leighton Pritchard, Paul R. J. Birch

    Research output: Contribution to journalArticle

    457 Citations (Scopus)

    Abstract

    Bacterial1, oomycete2 and fungal3 plant pathogens establish disease by translocation of effector proteins into host cells, where they may directly manipulate host innate immunity. In bacteria, translocation is through the type III secretion system1, but analogous processes for effector delivery are uncharacterized in fungi and oomycetes. Here we report functional analyses of two motifs, RXLR and EER, present in translocated oomycete effectors2. We use the Phytophthora infestans RXLR-EER-containing protein Avr3a as a reporter for translocation because it triggers RXLR-EER-independent hypersensitive cell death following recognition within plant cells that contain the R3a resistance protein4, 5. We show that Avr3a, with or without RXLR-EER motifs, is secreted from P. infestans biotrophic structures called haustoria, demonstrating that these motifs are not required for targeting to haustoria or for secretion. However, following replacement of Avr3a RXLR-EER motifs with alanine residues, singly or in combination, or with residues KMIK-DDK—representing a change that conserves physicochemical properties of the protein—P. infestans fails to deliver Avr3a or an Avr3a–GUS fusion protein into plant cells, demonstrating that these motifs are required for translocation. We show that RXLR-EER-encoding genes are transcriptionally upregulated during infection. Bioinformatic analysis identifies 425 potential genes encoding secreted RXLR-EER class proteins in the P. infestans genome. Identification of this class of proteins provides unparalleled opportunities to determine how oomycetes manipulate hosts to establish infection.
    Original languageEnglish
    Pages (from-to)115-118
    Number of pages4
    JournalNature
    Volume450
    Issue number7166
    DOIs
    Publication statusPublished - Nov 2007

    Fingerprint

    Oomycetes
    Phytophthora infestans
    host plants
    proteins
    secretion
    cells
    plant proteins
    bioinformatics
    plant pathogens
    infection
    alanine
    cell death
    physicochemical properties
    genes
    fungi
    genome
    bacteria

    Keywords

    • Phytophthora infestans
    • Pseudomonas-syringae
    • Potato infection
    • Downy mildew
    • Avirulence
    • Gene
    • Resistance
    • Secretion
    • Pathogen
    • Transcription

    Cite this

    Whisson, S. C., Boevink, P. C., Moleleki, L., Avrova, A. O., Morales, J. G., Gilroy, E. M., ... Birch, P. R. J. (2007). A translocation signal for delivery of oomycete effector proteins inside host plant cells. Nature, 450(7166), 115-118. https://doi.org/10.1038/nature06203
    Whisson, Stephen C. ; Boevink, Petra C. ; Moleleki, Lucy ; Avrova, Anna O. ; Morales, Juan G. ; Gilroy, Eleanor M. ; Armstrong, Miles R. ; Grouffaud, Severine ; West, Pieter van ; Chapman, Sean ; Hein, Ingo ; Toth, Ian K. ; Pritchard, Leighton ; Birch, Paul R. J. / A translocation signal for delivery of oomycete effector proteins inside host plant cells. In: Nature. 2007 ; Vol. 450, No. 7166. pp. 115-118.
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    abstract = "Bacterial1, oomycete2 and fungal3 plant pathogens establish disease by translocation of effector proteins into host cells, where they may directly manipulate host innate immunity. In bacteria, translocation is through the type III secretion system1, but analogous processes for effector delivery are uncharacterized in fungi and oomycetes. Here we report functional analyses of two motifs, RXLR and EER, present in translocated oomycete effectors2. We use the Phytophthora infestans RXLR-EER-containing protein Avr3a as a reporter for translocation because it triggers RXLR-EER-independent hypersensitive cell death following recognition within plant cells that contain the R3a resistance protein4, 5. We show that Avr3a, with or without RXLR-EER motifs, is secreted from P. infestans biotrophic structures called haustoria, demonstrating that these motifs are not required for targeting to haustoria or for secretion. However, following replacement of Avr3a RXLR-EER motifs with alanine residues, singly or in combination, or with residues KMIK-DDK—representing a change that conserves physicochemical properties of the protein—P. infestans fails to deliver Avr3a or an Avr3a–GUS fusion protein into plant cells, demonstrating that these motifs are required for translocation. We show that RXLR-EER-encoding genes are transcriptionally upregulated during infection. Bioinformatic analysis identifies 425 potential genes encoding secreted RXLR-EER class proteins in the P. infestans genome. Identification of this class of proteins provides unparalleled opportunities to determine how oomycetes manipulate hosts to establish infection.",
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    Whisson, SC, Boevink, PC, Moleleki, L, Avrova, AO, Morales, JG, Gilroy, EM, Armstrong, MR, Grouffaud, S, West, PV, Chapman, S, Hein, I, Toth, IK, Pritchard, L & Birch, PRJ 2007, 'A translocation signal for delivery of oomycete effector proteins inside host plant cells', Nature, vol. 450, no. 7166, pp. 115-118. https://doi.org/10.1038/nature06203

    A translocation signal for delivery of oomycete effector proteins inside host plant cells. / Whisson, Stephen C.; Boevink, Petra C.; Moleleki, Lucy; Avrova, Anna O.; Morales, Juan G.; Gilroy, Eleanor M.; Armstrong, Miles R.; Grouffaud, Severine; West, Pieter van; Chapman, Sean; Hein, Ingo; Toth, Ian K.; Pritchard, Leighton; Birch, Paul R. J.

    In: Nature, Vol. 450, No. 7166, 11.2007, p. 115-118.

    Research output: Contribution to journalArticle

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    AU - Whisson, Stephen C.

    AU - Boevink, Petra C.

    AU - Moleleki, Lucy

    AU - Avrova, Anna O.

    AU - Morales, Juan G.

    AU - Gilroy, Eleanor M.

    AU - Armstrong, Miles R.

    AU - Grouffaud, Severine

    AU - West, Pieter van

    AU - Chapman, Sean

    AU - Hein, Ingo

    AU - Toth, Ian K.

    AU - Pritchard, Leighton

    AU - Birch, Paul R. J.

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    N2 - Bacterial1, oomycete2 and fungal3 plant pathogens establish disease by translocation of effector proteins into host cells, where they may directly manipulate host innate immunity. In bacteria, translocation is through the type III secretion system1, but analogous processes for effector delivery are uncharacterized in fungi and oomycetes. Here we report functional analyses of two motifs, RXLR and EER, present in translocated oomycete effectors2. We use the Phytophthora infestans RXLR-EER-containing protein Avr3a as a reporter for translocation because it triggers RXLR-EER-independent hypersensitive cell death following recognition within plant cells that contain the R3a resistance protein4, 5. We show that Avr3a, with or without RXLR-EER motifs, is secreted from P. infestans biotrophic structures called haustoria, demonstrating that these motifs are not required for targeting to haustoria or for secretion. However, following replacement of Avr3a RXLR-EER motifs with alanine residues, singly or in combination, or with residues KMIK-DDK—representing a change that conserves physicochemical properties of the protein—P. infestans fails to deliver Avr3a or an Avr3a–GUS fusion protein into plant cells, demonstrating that these motifs are required for translocation. We show that RXLR-EER-encoding genes are transcriptionally upregulated during infection. Bioinformatic analysis identifies 425 potential genes encoding secreted RXLR-EER class proteins in the P. infestans genome. Identification of this class of proteins provides unparalleled opportunities to determine how oomycetes manipulate hosts to establish infection.

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    KW - Phytophthora infestans

    KW - Pseudomonas-syringae

    KW - Potato infection

    KW - Downy mildew

    KW - Avirulence

    KW - Gene

    KW - Resistance

    KW - Secretion

    KW - Pathogen

    KW - Transcription

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    Whisson SC, Boevink PC, Moleleki L, Avrova AO, Morales JG, Gilroy EM et al. A translocation signal for delivery of oomycete effector proteins inside host plant cells. Nature. 2007 Nov;450(7166):115-118. https://doi.org/10.1038/nature06203