Proteomics

Dataset Information

0

Exocyst Dynamics During Vesicle Tethering and Fusion


ABSTRACT: The exocyst is a conserved octameric complex that tethers exocytic vesicles to the plasma membrane prior to fusion. Exocyst assembly and delivery mechanisms remain unclear, especially in mammalian cells. Here we tagged multiple endogenous exocyst subunits with sfGFP or Halo using Cas9 gene-editing, to create single and double knock-in lines of mammary epithelial cells, and interrogated exocyst dynamics by high-speed imaging and correlation spectroscopy. We discovered that mammalian exocyst is comprised of tetrameric subcomplexes that can associate independently with vesicles and plasma membrane and are in dynamic equilibrium with octamer and monomers. Membrane arrival times are similar for subunits and vesicles, but with a small delay (~80msec) between subcomplexes. Departure of SEC3 occurs prior to fusion, whereas other subunits depart just after fusion. About 9 exocyst complexes are associated per vesicle. These data reveal the mammalian exocyst as a remarkably dynamic two-part complex and provide important insights into assembly/disassembly mechanisms.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Epithelial Cell

SUBMITTER: Syed Mukhtar Ahmed  

LAB HEAD: Ian G. Macara

PROVIDER: PXD011370 | Pride | 2018-12-05

REPOSITORIES: Pride

altmetric image

Publications

Exocyst dynamics during vesicle tethering and fusion.

Ahmed Syed Mukhtar SM   Nishida-Fukuda Hisayo H   Li Yuchong Y   McDonald W Hayes WH   Gradinaru Claudiu C CC   Macara Ian G IG  

Nature communications 20181203 1


The exocyst is a conserved octameric complex that tethers exocytic vesicles to the plasma membrane prior to fusion. Exocyst assembly and delivery mechanisms remain unclear, especially in mammalian cells. Here we tagged multiple endogenous exocyst subunits with sfGFP or Halo using Cas9 gene-editing, to create single and double knock-in lines of mammary epithelial cells, and interrogated exocyst dynamics by high-speed imaging and correlation spectroscopy. We discovered that mammalian exocyst is co  ...[more]

Similar Datasets

2002-12-22 | E-GEOD-92 | biostudies-arrayexpress
2019-01-31 | PXD009408 | Pride
2005-06-21 | E-GEOD-2830 | biostudies-arrayexpress
2022-10-14 | PXD031731 | Pride
2019-06-27 | PXD011212 | Pride
2023-08-09 | PXD043691 | Pride
2018-08-29 | PXD010336 | Pride
2022-10-19 | PXD033045 | Pride
2019-06-03 | PXD013835 | Pride
2017-02-14 | PXD002796 | Pride