TRIM62 promotes lipid raft-mediated entry of influenza A virus by regulating WASH localization

Abstract

Viruses employ diverse strategies to gain entry into cells to reach their replication sites. Influenza A virus (IAV), a respiratory pathogen, takes advantage of clathrin-dependent endocytosis or macropinocytosis for its internalization. Here, we report that TRIM62, a member of the tripartite motif (TRIM) protein family, is required for IAV entry, independent of its E3 ubiquitin ligase activity. We find that TRIM62 specifically functions in a clathrin-independent, lipid raft-mediated pathway, which IAV exploits to enter the endocytic network. Additionally, we reveal the involvement of the WASH complex and the retromer component VPS35 in this pathway beyond their canonical functions in endosomal sorting. We observe that a pool of WASH and VPS35 localize to the plasma membrane and associate with lipid rafts, in addition to their typical endosomal presence. While WASH subunits except FAM21 play a proviral role in IAV endocytosis and intracellular trafficking, VPS35 acts antagonistically. We show that TRIM62 counteracts the antiviral function of VPS35 by limiting FAM21-VPS35 interaction. By directly binding WASH in the cytosol, TRIM62 restricts its VPS35-mediated endosomal recruitment and thereby enhances its surface availability to facilitate IAV entry. Together, this study uncovers previously unrecognized roles of TRIM62 and endosomal sorting machinery in IAV entry, offering new antiviral targets.