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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. 3D modeling of AeCRN5 Cter sequence using AlphaFold2 predicted a structural homology with Restriction Endonuclease 5 (REase 5) family and identified a putative homodimerization of the effector. Co-immunoprecipitation analyses confirmed that AeCRN5 acts as a homodimer within the plant cells. A nucleic acid-protein interaction assay based on FRET-FLIM in N. benthamiana leaves revealed the RNA binding ability of AeCRN5 Cter domain. On-going experiments will help to evaluate a putative RNase activity of the effector. Alanine replacement of amino acids predicted to interact with RNA, abolished nuclear bodies accumulation of the AeCRN5 as well as effects on plant development, although the conservation of homodimerization of the effector. While RNA-binding capacity of AeCRN5 mutants are under investigation, 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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