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DataSet Summary

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

Dataset History

VersionDatetimeStatusChangeLog Entry
02018-10-16 06:28:03ID requested
12018-12-05 07:50:47announced
22019-04-03 04:32:50announcedUpdated publication reference for PubMed record(s): 30510181.

Publication List

  1. 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]

Keyword List

  1. submitter keyword: Vesicles, membrane fusions, CRISPR, exocyst

Contact List

    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:

Full Dataset Link List

  1. Dataset FTP location
  2. PRIDE project URI
Repository Record List

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