Uptake of oomycete RXLR effectors into host cells by clathrin-mediated endocytosis

• DOI: 10.1093/plcell/koad069
• Publication Date: 2023-03-13T15:47:17Z
• Authors: Haixia Wang; Shumei Wang; Wei Wang; Lin Xu; Lydia R J Welsh; Marek Gierlinski; Stephen C Whisson; Piers A Hemsley; Petra C Boevink; Paul R J Birch
• Keywords: Rab

Abstract Filamentous (oomycete and fungal) plant pathogens deliver cytoplasmic effector proteins into host cells to facilitate disease. How RXLR effectors from the potato late blight pathogen Phytophthora infestans enter is unknown. One possible route involves clathrin-mediated endocytosis (CME). Transient silencing of NbCHC, encoding clathrin heavy chain, or endosome marker gene NbAra6 a Rab GTPase in model Nicotiana benthamiana, attenuated P. infection reduced translocation fusions transgenic strains cells. By contrast, PP1c isoforms, susceptibility factors not required for endocytosis, but did attenuate uptake. Endosome enrichment by ultracentrifugation sucrose gradient fractionation revealed co-localization Pi04314-RFP with clathrin-coated vesicles. Immunopurification clathrin- NbAra6-associated vesicles during showed that AvrBlb1-RFP, apoplastic PiSCR74-RFP, were co-immunoprecipitated secreting these effectors. Tandem mass spectrometry analyses NbAra6-GFP associated endocytic alongside multiple effectors, including PiSCR74, which do Our data show uptake occurs via CME.