PXD011370

PXD011370 is an original dataset announced via ProteomeXchange.

Title	Exocyst Dynamics During Vesicle Tethering and Fusion
Description	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.
HostingRepository	PRIDE
AnnounceDate	2018-12-05
AnnouncementXML	Submission_2019-04-03_04:32:49.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD011370
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Syed Ahmed
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	Ammonia-loss; Dehydrated; Oxidation; Acetyl; Carbamidomethyl
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2018-10-16 06:28:03	ID requested
1	2018-12-05 07:50:47	announced
⏵ 2	2019-04-03 04:32:50	announced	Updated publication reference for PubMed record(s): 30510181.

Ahmed SM, Nishida-Fukuda H, Li Y, McDonald WH, Gradinaru CC, Macara IG, Exocyst dynamics during vesicle tethering and fusion. Nat Commun, 9(1):5140(2018) [pubmed]

submitter keyword: Vesicles, membrane fusions, CRISPR, exocyst

Ian G. Macara
contact affiliation	Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN, 37240, United States
contact email	ian.g.macara@vanderbilt.edu
lab head
Syed Ahmed
contact affiliation	Vanderbilt University School of Medicine
contact email	mukhtar06@gmail.com
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/12/PXD011370

PRIDE project URI

• PRIDE
  1. PXD011370
     1. Label: PRIDE project
     2. Name: Exocyst Dynamics During Vesicle Tethering and Fusion