Prion-like domains control plastid targeting of PEPSI effectors and PEPSI6-mediated modulation of DXR during root symbiosis

Abstract

Microbial effectors often act in the apoplast or cytosol, but how they reach host organelles during beneficial plant-fungal interactions remains poorly understood. Here, we identify a class of secreted prion-like effector proteins from the mutualistic root endophyte Serendipita indica, termed PEPSIs. These proteins contain prion-like domains (PrLDs) embedded within intrinsically disordered regions, and three tested PEPSIs require these domains for plastid localization. Focusing on PEPSI6, we show that plastid targeting depends on its PrLD and is associated with engagement of plastid import and proteostasis machinery. In plastids, PEPSI6 associates with 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR), promotes DXR accumulation, alters methylerythritol phosphate pathway metabolites, and enhances tolerance to the DXR inhibitor fosmidomycin. PEPSI6 also undergoes apoplastic C-terminal CAP-domain processing, and its colonization-promoting activity persists after PrLD deletion, indicating a plastid-targeting-independent function. Overall, this work identifies PrLDs as noncanonical plastid-targeting elements in PEPSI effectors and reveals DXR as a target of fungal manipulation during root symbiosis. HighlightsO_LIPrion-like domains control plastid targeting of selected PEPSI effectors. C_LIO_LIPEPSI6 associates with DXR and promotes its accumulation in plastids. C_LIO_LIPEPSI6 alters MEP-pathway metabolites and increases tolerance to fosmidomycin. C_LIO_LIApoplastic processing reveals a plastid-independent PEPSI6 activity during root symbiosis. C_LI