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Aphanomyces euteiches Crinkler effector interferes with RNA silencing
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International audience. Crinklers (CRN) effectors are widely distributed among oomycetes and numerous are known to target the host nucleus. Here we address the molecular function of AeCRN5 from the legume root pathogen Aphanomyces euteiches. As others CRNs oomycete effectors, AeCRN5 is a modular protein with a conserved Nterminus for plant targeting and a variable Cter domain with an unknown function. We showed that AeCRN5 exhibits dynamic nuclear localisation in plant cells, transiently accumulates in nuclear bodies, triggers cell death and induces major developmental defects when expressed in host root cells. Moreover, sequence analyses of AeCRN5 Cter revealed a structural homology with Restriction Endonuclease 5 (REase 5) family, including five amino acids highly conserved involved in the interaction with nucleic acids. We found that AeCRN5 acts as a homodimer and confirmed the nucleic acids affinity, especially for dsRNA, with in planta and in vitro experiments using wild type or mutated version of AeCRN5 by alanine replacement of amino acids predicted to interact with RNA/DNA. Interestingly, in planta expression of the mutated version of AeCRN5 revealed that nuclear bodies accumulation was abolished as well as effects on plant development, confirming the role of the conserved amino acids in the biological function of the protein. Finally, we found that AeCRN5 is able to interfere with plant RNA silencing mechanism using a heterologous system. Altogether, these data indicate that AeCRN5 acts through its Cter domain as a plant RNA silencing suppressor probably to facilitate pathogen infection
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