Organelle Tethering, Pore Formation and SNARE Compensation in the Late Endocytic Pathway

Overview

Journal J Cell Sci

Specialty Cell Biology

Date 2021 May 27

PMID 34042162

Citations 6

Authors

Luther J Davis

Nicholas A Bright

James R Edgar

Michael D J Parkinson

Lena Wartosch

Judith Mantell

Andrew A Peden

J Paul Luzio

Affiliations

Soon will be listed here.

Abstract

To provide insights into the kiss-and-run and full fusion events resulting in endocytic delivery to lysosomes, we investigated conditions causing increased tethering and pore formation between late endocytic organelles in HeLa cells. Knockout of the soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) VAMP7 and VAMP8 showed, by electron microscopy, the accumulation of tethered lysosome-associated membrane protein (LAMP)-carrier vesicles around multivesicular bodies, as well as the appearance of 'hourglass' profiles of late endocytic organelles attached by filamentous tethers, but did not prevent endocytic delivery to lysosomal hydrolases. Subsequent depletion of the SNARE YKT6 reduced this delivery, consistent with it compensating for the absence of VAMP7 and VAMP8. We also investigated filamentous tethering between multivesicular bodies and enlarged endolysosomes following depletion of charged multi-vesicular body protein 6 (CHMP6), and provide the first evidence that pore formation commences at the edge of tether arrays, with pore expansion required for full membrane fusion.

Citing Articles

Electron Tomography of Organelles and Vesicles in the Investigation of SNARE Function and Localization.

Bright N, Luzio J Methods Mol Biol. 2025; 2887():237-247.

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Four distinct cytoplasmic structures generate and release specific vesicles, thus opening the way to intercellular communication.

Racchetti G, Meldolesi J Extracell Vesicles Circ Nucl Acids. 2024; 4(1):44-58.

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Transient pores in hemifusion diaphragms.

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Reversible assembly and disassembly of V-ATPase during the lysosome regeneration cycle.

Sava I, Davis L, Gray S, Bright N, Luzio J Mol Biol Cell. 2024; 35(5):ar63.

PMID: 38446621 PMC: 11151095. DOI: 10.1091/mbc.E23-08-0322.

Loss of the HOPS complex disrupts early-to-late endosome transition, impairs endosomal recycling and induces accumulation of amphisomes.

van der Beek J, de Heus C, Sanza P, Liv N, Klumperman J Mol Biol Cell. 2024; 35(3):ar40.

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