Subunit connectivity, assembly determinants, and architecture of the yeast exocyst complex

Details

• Alternative Title:
  Nat Struct Mol Biol
• Personal Author:
  Heider, Margaret R. ; Gu, Mingyu ; Duffy, Caroline M. ; Mirza, Anne M. ; Marcotte, Laura L. ; Walls, Alexandra C. ; Farrall, Nicholas ; Hakhverdyan, Zhanna ; Field, Mark C. ; Rout, Michael P. ; Frost, Adam ; Munson, Mary
• Description:
  The exocyst is a hetero-octameric complex that has been proposed to serve as the tethering complex for exocytosis, although it remains poorly understood at the molecular level. Here, we purified endogenous exocyst complexes from Saccharomyces cerevisiae and showed that they are stable and consist of all eight subunits with equal stoichiometry. Using a combination of biochemical and auxin induced-degradation experiments in yeast, we mapped the subunit connectivity, identified two stable four-subunit modules within the octamer and demonstrated that several known exocyst-binding partners are not necessary for exocyst assembly and stability. Furthermore, we visualized the structure of the yeast complex by using negative-stain electron microscopy; our results indicate that the exocyst exists predominantly as a stable, octameric complex with an elongated architecture that suggests that the subunits are contiguous helical bundles packed together into a bundle of long rods.
• Subjects:
  Exocytosis Macromolecular Substances Microscopy, Electron, Transmission Protein Binding Protein Stability Protein Structure, Quaternary Saccharomyces Cerevisiae Vesicular Transport Proteins
• Source:
  Nat Struct Mol Biol. 23(1):59-66
• Pubmed ID:
  26656853
• Pubmed Central ID:
  PMC4752824
• Document Type:
  Journal Article
• Funding:
  GM068803/GM/NIGMS NIH HHSUnited States/ ; 1DP2GM110772/DP/NCCDPHP CDC HHSUnited States/ ; R01 GM068803/GM/NIGMS NIH HHSUnited States/ ; U54 GM103511/GM/NIGMS NIH HHSUnited States/ ; P41 GM109824/GM/NIGMS NIH HHSUnited States/ ; DP2 GM110772/GM/NIGMS NIH HHSUnited States/
• Volume:
  23
• Issue:
  1
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:5e03a068e91e8682010f253f33d550504c8e27f15cc27543cbfbae1352a84f96
• Download URL:
  https://stacks.cdc.gov/view/cdc/39147/cdc_39147_DS1.pdf
• File Type:
  [PDF - 1.58 MB ]