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

Margaret R. Heider, Mingyu Gu, Caroline M. Duffy, Anne M. Mirza, Laura L. Marcotte, Alexandra C. Walls, Nicholas Farrall, Zhanna Hakhverdyan, Mark C. Field, Michael P. Rout, Adam Frost, Mary Munson

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52 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)59-66
Number of pages8
JournalNature Structural & Molecular Biology
Volume23
Issue number1
Early online date14 Dec 2015
DOIs
Publication statusPublished - Jan 2016

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    Heider, M. R., Gu, M., Duffy, C. M., Mirza, A. M., Marcotte, L. L., Walls, A. C., Farrall, N., Hakhverdyan, Z., Field, M. C., Rout, M. P., Frost, A., & Munson, M. (2016). Subunit connectivity, assembly determinants and architecture of the yeast exocyst complex. Nature Structural & Molecular Biology, 23(1), 59-66. https://doi.org/10.1038/nsmb.3146