Endofin is required for HD-PTP and ESCRT-0 interdependent endosomal sorting of ubiquitinated transmembrane cargoes

Jalal M Kazan; Guillaume Desrochers; Claire E Martin; Hyeonju Jeong; Dmitri Kharitidi; Pirjo M Apaja; Ariel Roldan; Nicole St Denis; Anne-Claude Gingras; Gergely L Lukacs; Arnim Pause

doi:10.1016/j.isci.2021.103274

Endofin is required for HD-PTP and ESCRT-0 interdependent endosomal sorting of ubiquitinated transmembrane cargoes

iScience. 2021 Oct 14;24(11):103274. doi: 10.1016/j.isci.2021.103274. eCollection 2021 Nov 19.

Authors

Jalal M Kazan^{1

2}, Guillaume Desrochers^{1

2}, Claire E Martin³, Hyeonju Jeong^{1

2}, Dmitri Kharitidi^{1

2}, Pirjo M Apaja⁴, Ariel Roldan⁴, Nicole St Denis³, Anne-Claude Gingras^{3

5}, Gergely L Lukacs^{2

4}, Arnim Pause^{1

2}

Affiliations

¹ Goodman Cancer Research Center, McGill University, Montreal, QC H3A 1A3, Canada.
² Biochemistry Department, McGill University, Montreal, QC H3G 1Y6, Canada.
³ Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, Canada.
⁴ Physiology Department, McGill University, Montreal, QC H3G 1Y6, Canada.
⁵ Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.

Abstract

Internalized and ubiquitinated signaling receptors are silenced by their intraluminal budding into multivesicular bodies aided by the endosomal sorting complexes required for transport (ESCRT) machinery. HD-PTP, an ESCRT protein, forms complexes with ESCRT-0, -I and -III proteins, and binds to Endofin, a FYVE-domain protein confined to endosomes with poorly understood roles. Using proximity biotinylation, we showed that Endofin forms a complex with ESCRT constituents and Endofin depletion increased integrin α5-and EGF-receptor plasma membrane density and stability by hampering their lysosomal delivery. This coincided with sustained receptor signaling and increased cell migration. Complementation of Endofin- or HD-PTP-depleted cells with wild-type Endofin or HD-PTP, but not with mutants harboring impaired Endofin/HD-PTP association or cytosolic Endofin, restored EGFR lysosomal delivery. Endofin also promoted Hrs indirect interaction with HD-PTP. Jointly, our results indicate that Endofin is required for HD-PTP and ESCRT-0 interdependent sorting of ubiquitinated transmembrane cargoes to ensure efficient receptor desensitization and lysosomal delivery.

Keywords: Biological sciences; Cell biology; Molecular biology.