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  • Intracellular nanovesicles mediate α5β1 integrin trafficking during cell migration.

Intracellular nanovesicles mediate α5β1 integrin trafficking during cell migration.

The Journal of cell biology (2021-07-22)
Gabrielle Larocque, Daniel J Moore, Méghane Sittewelle, Cansu Kuey, Joseph H R Hetmanski, Penelope J La-Borde, Beverley J Wilson, Nicholas I Clarke, Patrick T Caswell, Stephen J Royle
ABSTRACT

Membrane traffic is an important regulator of cell migration through the endocytosis and recycling of cell surface receptors such as integrin heterodimers. Intracellular nanovesicles (INVs) are transport vesicles that are involved in multiple membrane trafficking steps, including the recycling pathway. The only known marker for INVs is tumor protein D54 (TPD54/TPD52L2), a member of the TPD52-like protein family. Overexpression of TPD52-like family proteins in cancer has been linked to poor prognosis and an aggressive metastatic phenotype, which suggests cell migration may be altered under these conditions. Here, we show that TPD54 directly binds membrane and associates with INVs via a conserved positively charged motif in its C terminus. We describe how other TPD52-like proteins are also associated with INVs, and we document the Rab GTPase complement of all INVs. Depletion of TPD52-like proteins inhibits cell migration and invasion, while their overexpression boosts motility. We show that inhibition of migration is likely due to altered recycling of α5β1 integrins in INVs.

MATERIALS
Product Number
Brand
Product Description

Roche
Anti-GFP, from mouse IgG1κ (clones 7.1 and 13.1)
Millipore
Anti-FLAG® M2 Magnetic Beads, affinity isolated antibody
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Anti-Integrin alpha5 (Preservative Free) Antibody, clone SNAKA51, clone SNAKA51, from mouse, purified by using protein G