Dataset Information

The endocytic TPLATE complex internalizes ubiquitinated plasma membrane cargo

ABSTRACT: Endocytosis controls the perception of stimuli by modulating protein abundance at the plasma membrane. In plants, clathrin-mediated endocytosis is the most prominent internalization pathway and relies on two multimeric adaptor complexes, the AP-2 and the TPLATE complex (TPC). Ubiquitination is a well-established modification triggering endocytosis of cargo proteins, but how this modification is recognized to initiate the endocytic event remains elusive. Here, we show that TASH3, one of the large subunits of TPC, recognizes ubiquitinated cargo at the plasma membrane via its SH3 domain-containing appendage. TASH3 lacking this evolutionary specific appendage modification allows TPC formation, but the plants show severely reduced endocytic densities, which correlates with reduced endocytic flux. Moreover, comparative plasma membrane proteomics identified differential accumulation of multiple ubiquitinated cargo proteins for which we confirm altered trafficking. Our findings position TPC as a key player for ubiquitinated cargo internalization, allowing future identification of target proteins under specific stress conditions.

INSTRUMENT(S): Q Exactive HF, Q Exactive

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Seedling

SUBMITTER: Dominique Eeckhout

LAB HEAD: Daniel Van Damme

PROVIDER: PXD035444 | Pride | 2023-03-11

REPOSITORIES: Pride

ACCESS DATA

Publications

The endocytic TPLATE complex internalizes ubiquitinated plasma membrane cargo.

Grones Peter P De Meyer Andreas A Pleskot Roman R Mylle Evelien E Kraus Michael M Vandorpe Michael M Yperman Klaas K Eeckhout Dominique D Dragwidge Jonathan Michael JM Jiang Qihang Q Nolf Jonah J Pavie Benjamin B De Jaeger Geert G De Rybel Bert B Van Damme Daniël D

Nature plants 20221201 12

Endocytosis controls the perception of stimuli by modulating protein abundance at the plasma membrane. In plants, clathrin-mediated endocytosis is the most prominent internalization pathway and relies on two multimeric adaptor complexes, the AP-2 and the TPLATE complex (TPC). Ubiquitination is a well-established modification triggering endocytosis of cargo proteins, but how this modification is recognized to initiate the endocytic event remains elusive. Here we show that TASH3, one of the large ...[more]

PMID: 36456802

Similar Datasets

Project description:The phylum Apicomplexa comprises an ancient group of early divergent eukaryotes, including some of the most deadly pathogens of medical and veterinary importance. Plasmodium species are responsible for malaria, which causes as many as 700,000 deaths per year, while Toxoplasma gondii chronically infects up to 30% of the human population, with immunocompromised patients and pregnant women at risk for adverse outcomes, such as toxoplasmic encephalitis and spontaneous abortion, respectively. T. gondii is considered a model system not only for its pathogenic relatives but also for intracellular parasitism and infection biology in general. T. gondii has common eukaryotic organelles, including the nucleus, endoplasmic reticulum (ER), and a single Golgi stack, but also specific secretory organelles named dense granules, micronemes, and rhoptries that contain parasite-derived factors required for host infection. Rhoptries and micronemes are formed de novo during parasite replication, and this process requires significant protein and lipid trafficking through the secretory pathway. The trafficking mechanisms employed by T. gondii retain several typical eukaryote components as well as evolving divergent features. Protein trafficking of this parasite is mediated by entry into a canonical ER followed by vesicle packaging through a single Golgi complex. Post-Golgi protein sorting to specific organelles requires the function of dynamin-related protein B, which is involved in fission events. Downstream Rab-GTPases function throughout the parasite secretory pathway. T. gondii soluble N-ethylmaleimide-sensitive-factor attachment protein receptor (SNARE) proteins in docking and fusion at target membranes have also been described. However, unlike in mammalian cells, T. gondii ER is reduced so that the nuclear envelope itself contributes to a substantial proportion of its total volume. Whereas in mammalian cells hundreds of Golgi stacks occupy the perinuclear area, the Golgi apparatus is limited to a single discrete structure in T.gondii. The post-Golgi system also named the endosome-like compartment (ELC) is involved in the trafficking of microneme proteins. The ELC is decorated by the small GTPases, Rab5 and Rab7, which are typically associated with the endosomal system. Nevertheless, classical endocytosis has not yet been validated in T. gondii. This parasite has no lysosomes; rather the parasite harbors acidic vesicles that were thought to be precursors of the rhoptry organelles. The parasite lacks most components of endosomal sorting complexes (ESCRT), which are known for their roles in forming multivesicular bodies that deliver ubiquitinated membrane proteins and lipids to lysosomes for degradation. The machinery required for caveogenesis and caveola-dependent invaginations have not yet been identified in the parasite. Furthermore, while evidence of conventional clathrin-dependent endocytosis by T.gondii is lacking, clathrin is present exclusively in post-Golgi compartments where its function is restricted to post-Golgi trafficking, and the uptake of cytosol proteins by the tachyzoites of T. gondii has recently been described using an endocytosis assay. However, the mechanisms underlying the events of this unconventional endocytosis in the parasite remain to be determined. Clearly, the secretory pathway of T. gondii can be considered a stripped-down version of the more complex trafficking machinery that characterizes higher eukaryotes. Despite this minimal trafficking machinery, the parasites actively rely on a membrane vesicle formation and transport during its intracellular lifecycle; however, to date, comparatively little is known about the mechanisms involved in trafficking pathways in T. gondii. We previously reported a T. gondii sortilin-like receptor (TgSORTLR) that regulates protein transport and is essential for apical secretory organelle biogenesis and host infection17. Moreover, the C-terminal tail of TgSORTLR was shown to be involved in recruiting many cytosolic cargo proteins including two homologues of the core retromer components, Vps26 and Vps35, which are known to regulate retrograde transport from endosomes to the trans-Golgi Network (TGN) in yeast and mammals. Here, we report that a singular architecture with a trimer Vps35-Vps26-Vps29 core complex acts as the major cargo endosomal recycling machinery and is required for parasite integrity and more specifically for secretory organelle biogenesis and maintenance of a multiple ligand-binding transporter at the T. gondii membrane. Our findings provide strong evidence that the unconventional TgSORTLR-containing endosome-like compartment is involved in distinct mechanisms for the delivery of major retromer-dependent cargo. In this current work, we demonstrate a definitive role for the endocytic recycling pathway in T. gondii pathogenesis.

Dataset Information

The endocytic TPLATE complex internalizes ubiquitinated plasma membrane cargo

Publications

The endocytic TPLATE complex internalizes ubiquitinated plasma membrane cargo.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure