Fusion of Madin-Darby canine kidney cells by HVJ (Sendai virus): absence of direct association of virus particles with the site of membrane fusion.
The process of cell fusion of Madin-Darby canine kidney (MDCK) cells by HVJ (Sendai virus) was investigated to determine whether the HVJ particles were directly associated with the site of membrane fusion. Confluent monolayer cultures of MDCK cells are sealed together by tight junctions on the apices of their lateral membranes, so added virus particles can be adsorbed only to the apical surfaces of the cells. After incubation with HVJ at 37 degrees C for 30 min, the cells still appeared mononucleate and unfused by light microscopy, but electron microscopic examination showed that fusion at the lateral membranes had occurred below the tight junctions. Furthermore, when fluorescein isothiocyanate (FITC)-labeled macromolecules, which cannot pass across the gap junctions, were injected into the cells at this stage, labeled macromolecules were found to diffuse into the adjacent cells. These findings strongly suggest that cell fusion was initiated in the lateral membrane, a region distinct from the site of adsorbed HVJ particles. Thus, the virus particles were not directly associated with the fusion site, but induced fusion of the lateral membranes indirectly.
Kim
J
J
Institute of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University, Japan.
Adachi
T
T
Yoneda
Y
Y
Okada
Y
Y
D000818:Animals; D002459:Cell Fusion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000382:microbiology / Q000648:ultrastructure; D004058:Diffusion; D004285:Dogs; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D005452:Fluoresceins / Q000378:metabolism; D007365:Intercellular Junctions / Q000648:ultrastructure; D008561:Membrane Fusion; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D010222:Parainfluenza Virus 1, Human / Q000502:physiology*; D012710:Serum Albumin, Bovine / Q000378:metabolism
D005452:Fluoresceins; C039217:fluorescein isothiocyanate bovine serum albumin; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate