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The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking
Published on: February 12, 2022
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COMMD5/HCaRG Hooks Endosomes on Cytoskeleton and Coordinates EGFR Trafficking
Carole G Campion1, Kossay Zaoui2, Thomas Verissimo1
1Centre de Recherche, Centre Hospitalier de l'Université de Montréal (CRCHUM), Montréal, QC H2X 0A9, Canada; Département de Médecine, Université de Montréal, Montréal, QC H3T 1J4, Canada.
Cell Reports
|July 19, 2018
Summary
COMMD5 protein stabilizes the cytoskeleton and is essential for endosomal trafficking, including epidermal growth factor receptor (EGFR) recycling. Its dysfunction impacts cell structure and EGFR transport, crucial for tissue repair and cancer.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- COMMD5/HCaRG is implicated in tissue repair and its reduced expression correlates with tumorigenicity.
- COMMD5 regulates cell growth, migration, and differentiation, and previously shown to inhibit renal carcinoma cell proliferation.
- COMMD5's role in cytoskeletal stability and endosomal trafficking was not fully elucidated.
Purpose of the Study:
- To elucidate the molecular mechanism by which COMMD5 influences cytoskeletal stability.
- To investigate COMMD5's role in endosomal trafficking and epidermal growth factor receptor (EGFR) recycling.
- To establish COMMD5's function as an adaptor protein coordinating endosomal transport and cellular activity.
Main Methods:
- COMMD5 silencing to observe cytoskeletal re-organization (actin and microtubule networks).
- Co-immunoprecipitation and binding assays to identify COMMD5 N- and C-terminal interactions with Rab5 and cytoskeletal components.
- Analysis of endosome transport, including EGFR recycling, in COMMD5-silenced cells.
Main Results:
- COMMD5 silencing caused significant re-organization of actin and microtubule networks, highlighting its role in cytoskeletal stability.
- The N terminus of COMMD5 binds to endosomal Rab5, while the C terminus interacts with cytoskeletal scaffolding.
- COMMD5 facilitates long-range endosome transport, crucial for EGFR recycling and vesicle scission into sorting endosomes.
Conclusions:
- COMMD5 is essential for maintaining cytoskeletal integrity.
- COMMD5 acts as a molecular adaptor, linking endosomal trafficking (via Rab5) with cytoskeletal dynamics.
- COMMD5 plays a critical role in EGFR transport and activity, impacting cellular functions relevant to tissue repair and cancer.
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