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Exocyst stimulates each step of exocytic SNARE complex assembly and vesicle fusion

View ORCID ProfileChanwoo Lee, View ORCID ProfileDante Lepore, View ORCID ProfileMary Munson, View ORCID ProfileTae-Young Yoon
doi: https://doi.org/10.1101/2022.01.16.476540
Chanwoo Lee
1School of Biological Sciences and Institute for Molecular Biology and Genetics Seoul National University, Seoul 08826, South Korea
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Dante Lepore
2Department of Biochemistry and Molecular Pharmacology University of Massachusetts Medical School, 364 Plantation Street, Worcester, MA 01605 USA
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Mary Munson
2Department of Biochemistry and Molecular Pharmacology University of Massachusetts Medical School, 364 Plantation Street, Worcester, MA 01605 USA
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  • For correspondence: mary.munson@umassmed.edu tyyoon@snu.ac.kr
Tae-Young Yoon
1School of Biological Sciences and Institute for Molecular Biology and Genetics Seoul National University, Seoul 08826, South Korea
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  • For correspondence: mary.munson@umassmed.edu tyyoon@snu.ac.kr
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Abstract

The exocyst is a large multisubunit tethering complex essential for targeting and fusion of secretory vesicles in eukaryotic cells. Although the assembled exocyst complex is proposed to tether vesicles to the plasma membrane and activate the SNARE proteins for membrane fusion, only little is known about the key biochemical steps that exocyst stimulates in the course of SNARE complex assembly, a critical question defining the essential molecular role of the exocyst complex. Here, we use a combination of single molecule and bulk fluorescence assays with purified octameric yeast exocyst complexes to examine the role of exocyst in a reconstituted SNARE assembly and fusion system. We show that the exocyst complex simulates multiple steps spanning from SNARE protein activation to ternary complex assembly, rather than affecting only a specific subset of steps. We also observed that the exocyst has important downstream roles in driving membrane fusion, up to full content mixing of vesicle lumens. Our results suggest that the exocyst complex provides extensive chaperoning functions for the entire process of SNARE complex assembly, presumably using its multi-faceted structure provided by the eight subunits.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 17, 2022.
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Exocyst stimulates each step of exocytic SNARE complex assembly and vesicle fusion
Chanwoo Lee, Dante Lepore, Mary Munson, Tae-Young Yoon
bioRxiv 2022.01.16.476540; doi: https://doi.org/10.1101/2022.01.16.476540
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Exocyst stimulates each step of exocytic SNARE complex assembly and vesicle fusion
Chanwoo Lee, Dante Lepore, Mary Munson, Tae-Young Yoon
bioRxiv 2022.01.16.476540; doi: https://doi.org/10.1101/2022.01.16.476540

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