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The In Vivo Architecture of the Exocyst Provides Structural Basis for Exocytosis
- Picco, Andrea, Irastorza-Azcarate, Ibai, Specht, Tanja, Böke, Dominik, Pazos, Irene, Rivier-Cordey, Anne-Sophie, Devos, Damien P., Kaksonen, Marko, Gallego, Oriol
- Cell 2017 v.168 no.3 pp. 400-412.e18
- direct contact, exocytosis, fluorescence microscopy, plasma membrane, secretory granules, yeasts
- The structural characterization of protein complexes in their native environment is challenging but crucial for understanding the mechanisms that mediate cellular processes. We developed an integrative approach to reconstruct the 3D architecture of protein complexes in vivo. We applied this approach to the exocyst, a hetero-octameric complex of unknown structure that is thought to tether secretory vesicles during exocytosis with a poorly understood mechanism. We engineered yeast cells to anchor the exocyst on defined landmarks and determined the position of its subunit termini at nanometer precision using fluorescence microscopy. We then integrated these positions with the structural properties of the subunits to reconstruct the exocyst together with a vesicle bound to it. The exocyst has an open hand conformation made of rod-shaped subunits that are interlaced in the core. The exocyst architecture explains how the complex can tether secretory vesicles, placing them in direct contact with the plasma membrane.