Dissecting the cell entry pathway of dengue virus by single-particle tracking in living cells.
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.
van der Schaar
Hilde M
HM
Department of Medical Microbiology, Molecular Virology Section, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Rust
Michael J
MJ
Chen
Chen
C
van der Ende-Metselaar
Heidi
H
Wilschut
Jan
J
Zhuang
Xiaowei
X
Smit
Jolanda M
JM
D000330:Aedes; D000818:Animals; D002478:Cells, Cultured; D022163:Clathrin-Coated Vesicles / Q000378:metabolism / Q000821:virology; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000502:physiology*; D004058:Diffusion; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D015398:Signal Transduction / Q000502:physiology; D022161:Transport Vesicles / Q000473:pathology / Q000648:ultrastructure*; D014771:Virion / Q000378:metabolism*; D053586:Virus Internalization; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
D000090802:rab7 GTP-Binding Proteins; C000716463:rab7 GTP-binding proteins, human; D020691:rab GTP-Binding Proteins