PXD040285

PXD040285 is an original dataset announced via ProteomeXchange.

Title	Uptake of oomycete RXLR effectors into host cells by clathrin-mediated endocytosis
Description	Filamentous (oomycete and fungal) plant pathogens can deliver cytoplasmic effector proteins into host cells to facilitate disease. How RXLR effectors from the potato late blight pathogen Phytophthora infestans enter host cells is unknown. One possible route involves clathrin-mediated endocytosis (CME). Transient silencing of NbCHC, encoding clathrin heavy chain, or endosome marker NbAra6 in the model host Nicotiana benthamiana, attenuated P. infestans infection and reduced translocation of RXLR effector fusions from transgenic pathogen lines into host cells. In contrast, silencing PP1c isoforms, susceptibility factors that are not required for endocytosis, reduced infection but did not attenuate RXLR effector uptake. Endosome enrichment by ultracentrifugation and sucrose gradient fractionation revealed co-localisation of RXLR effector Pi04314-RFP with clathrin-coated vesicles. Immunopurification of clathrin- and NbAra6-associated vesicles during infection revealed that RXLR effectors Pi04314-RFP and AvrBlb1-RFP, but not apoplastic effector PiSCR74-RFP, were co-immunoprecipitated during infection with pathogen lines secreting these effectors. MS/MS analyses of proteins co-immunoprecipitated with NbAra6-GFP during infection revealed enrichment of host proteins associated with endocytic vesicles alongside multiple pathogen RXLR effectors, but not apoplastic effectors, including PiSCR74, which do not enter host cells. Our data show that uptake of P. infestans RXLR effectors into plant cells occurs via CME.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:03:47.117.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Haixia Wang
SpeciesList	scientific name: Phytophthora infestans T30-4; NCBI TaxID: 403677; scientific name: Nicotiana benthamiana; NCBI TaxID: 4100;
ModificationList	acetylated residue; monohydroxylated residue; deaminated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-02-20 07:13:11	ID requested
1	2023-07-20 12:16:44	announced
⏵ 2	2023-11-14 09:03:49	announced	2023-11-14: Updated project metadata.

Wang H, Wang S, Wang W, Xu L, Welsh LRJ, Gierlinski M, Whisson SC, Hemsley PA, Boevink PC, Birch PRJ, Uptake of oomycete RXLR effectors into host cells by clathrin-mediated endocytosis. Plant Cell, 35(7):2504-2526(2023) [pubmed]

submitter keyword: oomycete

RXLR effectors

clathrin mediated endocytosis

Paul Birch
contact affiliation	University of Dundee
contact email	P.Birch@dundee.ac.uk
lab head
Haixia Wang
contact affiliation	University of Dundee
contact email	hzjwang@dundee.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD040285
     1. Label: PRIDE project
     2. Name: Uptake of oomycete RXLR effectors into host cells by clathrin-mediated endocytosis