Dissecting the Cell Entry Pathway of Dengue Virus by Single-Particle Tracking in Living Cells

TLDR

Dengue virus is an enveloped RNA virus that causes the most common arthropod‑borne infection worldwide, yet the cellular entry mechanism remains unclear. The study seeks to elucidate DENV cell entry, endocytic trafficking, and fusion behavior. Live‑cell imaging and single‑virus tracking were used to observe DENV movement and interactions during entry. DENV enters cells exclusively via clathrin‑mediated endocytosis, diffusing along the surface before capture by clathrin‑coated pits, then traffics to Rab5‑positive endosomes that mature into late endosomes through Rab7 acquisition and Rab5 loss, where fusion with the endosomal membrane occurs.

Abstract

Dengue virus (DENV) is an enveloped RNA virus that causes the most common arthropod-borne infection worldwide. The mechanism by which DENV infects the host cell remains unclear. In this work, we used live-cell imaging and single-virus tracking to investigate the cell entry, endocytic trafficking, and fusion behavior of DENV. Simultaneous tracking of DENV particles and various endocytic markers revealed that DENV enters cells exclusively via clathrin-mediated endocytosis. The virus particles move along the cell surface in a diffusive manner before being captured by a pre-existing clathrin-coated pit. Upon clathrin-mediated entry, DENV particles are transported to Rab5-positive endosomes, which subsequently mature into late endosomes through acquisition of Rab7 and loss of Rab5. Fusion of the viral membrane with the endosomal membrane was primarily detected in late endosomal compartments.