The TPLATE subunit is essential for structural assembly of the endocytic TSET complex

Abstract

All eukaryotic cells rely on endocytosis to regulate the plasma membrane proteome and lipidome. Most eukaryotic groups, with the exception of fungi and animals, have retained the evolutionary ancient TSET complex as a regulator of endocytosis. Despite the presence of similar building blocks in TSET, compared to other coatomer complexes, structural insight into this adaptor complex is lacking. Here, we elucidate the molecular architecture of the octameric plant TSET complex (TPLATE complex/TPC) using an integrative structural approach. This allowed us to describe a plant-specific connection between the TML subunit and the AtEH/Pan1 proteins and show a direct interaction between the complex and the plasma membrane without the need for any additional protein factors. Furthermore, we identify the appendage of TPLATE as crucial for complex assembly. Structural elucidation of this ancient adaptor complex vastly advances our functional as well as evolutionary insight into the process of endocytosis.

Visualizing protein–protein interactions in plants by rapamycin-dependent delocalization

“…The TPLATE complex core consists of four subunits: TPLATE, TML, LOLITA, and TASH3. Interactions between particular subunits have been demonstrated via different PPI methods (Gadeyne et al, 2014;Hirst et al, 2014;Yperman et al, 2020).…”

Visualizing protein-protein interactions in plants by rapamycin-dependent delocalization

“…The TPLATE complex core consists of four subunits: TPLATE, TML, LOLITA, and TASH3. Interactions between particular subunits have been demonstrated via different PPI methods (Gadeyne et al, 2014; Hirst et al, 2014; Yperman et al, 2020). We tested the TPLATE-TML interaction, as previously revealed via BiFC and Co-IP (Gadeyne et al, 2014), using KSP.…”