Dataset Information

Host-interactor screens of Phytophthora infestans RXLR proteins

ABSTRACT: Pathogens modulate host cell structure and function by secreting effectors into host tissues. Effectors generally function by associating with host molecules and co-opting or perturbing their activities. This study aimed at identifying the plant proteins associated with the RXLR class of host-translocated effectors of the potato blight pathogen Phytophthora infestans in order to identify the host processes targeted by these effectors. We carried out an in planta screen by transiently expressing P. infestans RXLR effectors in Nicotiana benthamiana leaves followed by co-immunoprecipitation (co-IP) and liquid chromatography tandem mass spectrometry (LC-MS/MS). This generated an effector-host protein interactome matrix of 59 P. infestans RXLR effectors x 586 N. benthamiana proteins. Classification of the host interactors into putative functional categories revealed over 35 biological processes that are candidate effector-targeted pathways. We further characterized the PexRD12/31 family of RXLR-WY effectors, which associate and co-localize with components of vesicle trafficking machinery. One member of this family, PexRD31, increased the number of GFP-2xFYVE labeled vesicles in N. benthamiana cells. We also noted an accumulation of FYVE vesicles in areas of N. benthamiana leaves colonized by P. infestans indicating that this pathogen may enhance endosomal trafficking. We hope that the interactome dataset we generated will serve as a useful community resource for functional studies of P. infestans effectors.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Phytophthora Infestans Nicotiana Benthamiana

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Jan Sklenar

LAB HEAD: Jan Sklenar

PROVIDER: PXD020751 | Pride | 2020-09-23

REPOSITORIES: Pride

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Publications

Host-interactor screens of Phytophthora infestans RXLR proteins reveal vesicle trafficking as a major effector-targeted process.

Petre Benjamin B Contreras Mauricio P MP Bozkurt Tolga O TO Schattat Martin H MH Sklenar Jan J Schornack Sebastian S Abd-El-Haliem Ahmed A Castells-Graells Roger R Lozano-Durán Rosa R Dagdas Yasin F YF Menke Frank L H FLH Jones Alexandra M E AME Vossen Jack H JH Robatzek Silke S Kamoun Sophien S Win Joe J

The Plant cell 20210701 5

Pathogens modulate plant cell structure and function by secreting effectors into host tissues. Effectors typically function by associating with host molecules and modulating their activities. This study aimed to identify the host processes targeted by the RXLR class of host-translocated effectors of the potato blight pathogen Phytophthora infestans. To this end, we performed an in planta protein-protein interaction screen by transiently expressing P. infestans RXLR effectors in Nicotiana bentham ...[more]

PMID: 33677602

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Project description:Intracellular bacterial pathogens inject a cocktail of effector proteins into host cells to hijack diverse cellular processes and promote their survival and proliferation. Salmonella enterica serovar Typhimurium (STm) uses two Type 3 secretion system (T3SS) to deliver a suite of effector proteins into host cells, however, knowledge of their host-targets remains sparse. To systematically map STm effector-host protein-protein interactions (PPIs) during active infection, we generated a library of 32 STm strains expressing affinity-tagged effector proteins from their endogenous chromosomal loci. Tagged-effector strains were then used to infect macrophages or epithelial cells, and PPIs were detected by Affinity-Purification Quantitative Mass-Spectrometry (AP-QMS) under both native and cross-linking conditions. We recovered 25 previously described effector-host PPIs, along with 421 novel interactions across the two cell lines tested, as well as several effector-effector PPIs. While effectors converged on specific host cellular processes, which varied across cell types, most had multiple host targets. Using reciprocal co-immunoprecipitations, we could validated 13 out of 22 new PPIs, demonstrating an accuracy of at least 59% for the AP-QMS approach. To illustrate the utility of this resource for discovering novel infection biology, we first showed that SseJ and SseL cooperate to control cholesterol transport at the Salmonella Containing Vacuole (SCV) via the Niemann-Pick C1 protein (NPC1). Second, we uncovered that PipB directly binds and recruits the organelle contact site protein PDZD8 to the SCV. Third, we elucidated a novel mechanism whereby the effector kinase SteC promotes host actin bundling, by directly interacting and phosphorylating formin-like proteins. Overall, we provide a systems-wide host-bacterial pathogen physical interactome resource, to our knowledge the first in an infection context, with broad implications for effector function and cooperation.

Dataset Information

Host-interactor screens of Phytophthora infestans RXLR proteins

Publications

Host-interactor screens of Phytophthora infestans RXLR proteins reveal vesicle trafficking as a major effector-targeted process.

Similar Datasets

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