SUMOylation of EHD3 Modulates Tubulation of the Endocytic Recycling Compartment

Abstract

Endocytosis defines the entry of molecules or macromolecules through the plasma membrane as well as membrane trafficking in the cell. It depends on a large number of proteins that undergo protein-protein and protein-phospholipid interactions. EH Domain containing (EHDs) proteins formulate a family, whose members participate in different stages of endocytosis. Of the four mammalian EHDs (EHD1-EHD4) EHD1 and EHD3 control traffic to the endocytic recycling compartment (ERC) and from the ERC to the plasma membrane, while EHD2 modulates internalization. Recently, we have shown that EHD2 undergoes SUMOylation, which facilitates its exit from the nucleus, where it serves as a co-repressor. In the present study, we tested whether EHD3 undergoes SUMOylation and what is its role in endocytic recycling. We show, both in-vitro and in cell culture, that EHD3 undergoes SUMOylation. Localization of EHD3 to the tubular structures of the ERC depends on its SUMOylation on lysines 315 and 511. Absence of SUMOylation of EHD3 has no effect on its dimerization, an important factor in membrane localization of EHD3, but has a dominant negative effect on its appearance in tubular ERC structures. Non-SUMOylated EHD3 delays transferrin recycling from the ERC to the cell surface. Our findings indicate that SUMOylation of EHD3 is involved in tubulation of the ERC membranes, which is important for efficient recycling.

A semi-supervised Bayesian approach for simultaneous protein sub-cellular localisation assignment and novelty detection

“…We observe a mixed steady-state potential localisation to the endosome and PM for EHD3 (gure 6, panel d). Our results are in agreement with previously published studies which have reported that EHD3 is resident in the early endosomes and recycling endosomes at steady state (Naslavsky et al, 2006(Naslavsky et al, , 2009George et al, 2007;Cabasso et al, 2015), and our PM localisation-related observation is supported by the HPA Cell Atlas-derived annotation for this protein (gure 6, panel b).…”

“…We observe a mixed steady-state potential localisation to the endosome and PM for EHD3 (gure 6, panel d). This is in agreement with EHD3's role in recycling endosome-to-PM transport (Naslavsky et al, 2006(Naslavsky et al, , 2009George et al, 2007;Cabasso et al, 2015;Henmi et al, 2016).…”

“…This protein promotes the biogenesis and stabilisation of tubular recycling endosomes by inducing early endosome membrane bending and tubulation (Bahl et al, 2016;Henmi et al, 2016). Additionally, EHD3 is essential for early endosome-to-recycling endosome transport, retrograde early endosome-to-Golgi trac, Golgi apparatus morphology maintenance, and recycling endosome-to-PM transport (Naslavsky et al, 2006(Naslavsky et al, , 2009George et al, 2007;Cabasso et al, 2015;Henmi et al, 2016). It plays an important role in the recycling of cell surface receptors and the biosynthetic transport of lysosome proteins (Naslavsky et al, 2006(Naslavsky et al, , 2009George et al, 2007;Cabasso et al, 2015).…”

A semi-supervised Bayesian approach for simultaneous protein sub-cellular localisation assignment and novelty detection

“…We observe a mixed steady-state potential localisation to the endosome and PM for EHD3 ( Fig 6D). This is in agreement with EHD3's role in recycling endosome-to-PM transport [80][81][82][83][84].…”

Posttranslational Modifications of Smurfs: Emerging Regulation in Cancer

“…Like Nedd8, SUMO is also a ubiquitin-like molecule, and at this point, the sumoylation process is understood as similar to that of ubiquitination ( 90 ). After being sumoylated, protein stability, cellular distribution, activity, and interactions can be governed ( 91 – 94 ). In mammary epithelial cell-derived acini, sumoylation of Smurf2 by the SUMO E3 ligase (PIAS3) at Lys26 and Lys369 enhances Smurf2 stability and ubiquitin ligase activity ( 54 ).…”