MassIVE MSV000094520

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Molecular Mechanisms of Stress Granule Disassembly Revealed by Chemogenetic Microenvironment Mapping (MicroMap) in Living Cells

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Description

Phase-separated condensates are membrane-less intracellular structures comprised of dynamic protein interactions that organize essential biological processes. Understanding the composition and dynamics of these organelles advances our knowledge of cellular behaviors and disease pathologies related to granule dysregulation. In this study, we apply microenvironment mapping (MicroMap) with a novel HaloTag-based platform (HaloMap) to characterize intracellular stress granule dynamics in living cells. After validating the robustness and sensitivity of this approach, we then profile the stress granule proteome throughout the formation and disassembly, and under pharmacological perturbation. These experiments reveal several novel ubiquitin-related modulators, including the HECT-type E3 ligases ITCH and NEDD4L, as well as the ubiquitin receptor TOLLIP, as key mediators of granule disassembly. In addition, we identify an autophagy-related pathway that promotes granule clearance. Collectively, this work establishes a general photoproximity labeling approach for unraveling intracellular protein interactomes and uncovers previously unexplored regulatory mechanisms of stress granule dynamics. [doi:10.25345/C5707X05X] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Stress granules ; Proximity labeling ; Granule disassembly ; MicroMap

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Principal Investigators:
(in alphabetical order) 		David MacMillan, Princeton, USA
Submitting User: RoderickPan
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