Global organelle profiling reveals subcellular localization and remodeling at proteome scale

Area of Science:

• Cell Biology
• Proteomics
• Systems Biology

Background:

• Understanding the precise location and dynamics of proteins within cells is crucial for deciphering cellular functions and responses.
• Existing methods often lack the resolution or scope to comprehensively map the entire proteome's subcellular distribution and its dynamic changes.

Purpose of the Study:

• To develop and apply a high-resolution strategy for mapping the subcellular localization of thousands of human proteins.
• To analyze proteome remodeling, particularly spatial distribution changes, during cellular perturbations like viral infection.
• To investigate the role of ferroptosis in human coronavirus OC43 (HCoV-OC43) infection.

Main Methods:

• Organelle immunocapture coupled with mass spectrometry (MS) to isolate and identify proteins within specific cellular compartments.
• Graph-based analysis to assign subcellular localization to over 7,600 proteins and define spatial networks.
• Proteomic profiling of cells before and after HCoV-OC43 infection to identify changes in protein abundance and localization.

Main Results:

• Successfully mapped the subcellular localization of over 7,600 human proteins, revealing extensive spatial networks and inter-compartmental connections.
• Demonstrated that viral infection (HCoV-OC43) induces significant protein remodeling primarily through changes in spatial distribution, not just abundance.
• Identified a critical role for ferroptosis in the cellular response to HCoV-OC43 infection.

Conclusions:

• Proteome-wide analysis of subcellular remodeling provides critical insights into cellular responses to infection.
• Spatial protein redistribution is a key mechanism regulating cellular states during viral pathogenesis.
• Ferroptosis is an essential component of the host defense against HCoV-OC43.