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Pucadyil Lab

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Discovering and analyzing functions of membrane fission proteins

Formation of organelles requires the limiting membrane to be constricted to nanometer dimensions before undergoing fission. Since organelles are of much larger dimensions, fission necessitates a topological transformation of organelles into a highly curved tube-like intermediate. Using Supported Membrane Templates (SMrT) that represent an array membrane nanotubes (see schematic below and Dar, Kamerkar and Pucadyil (2017) Nature Protocols for more details) and display a wide range of curvatures, mimicking intermediates in the process of fission, we screen tissue lysates for fission activity and identify the sets of proteins that contribute to such activity. We then purify and reconstitute their functions to better understand how they work. The hits that emerge from our screen are tested for cellular function by making knock-out models and analysing morphological changes to various membrane compartments.

SMrT templates.png
Dynamin fission (5% PIP2 cyan).gif

Schematic of formation of SMrTs and a movie showing fission of nanotubes seen upon flowing a fission protein.

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Workflow of the discovery process.

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