Interaction of rotavirus particles with liposomes.
We have studied the interactions of purified viral particles with liposomes as a model to understand the mechanism of entry of rotavirus into the cell. Liposomes, made from pure as well as mixed lipids, that contained encapsulated self-quenching concentrations of the fluorophore carboxyfluorescein (CF) were used. Rotavirus-liposome interactions were studied from the fluorescence dequenching of CF resulting from its release to the bulk solution. Purified infectious double-shelled virus particles induced a concentration- and temperature-dependent release of CF. The rate and extent of CF release was maximum between pH 7.3 and 7.6. The removal of outer structural proteins VP4 and VP7 from virus, which results in the formation of single-shelled particles, prevented virus interaction with liposomes. Rotavirus particles with uncleaved VP4 did not interact with liposomes, but treatment in situ of these particles with trypsin restored the interaction with the liposomes and resulted in CF dequenching. Our data support the view that rotavirus enters the cell through direct penetration of the plasma membrane. In contrast, adenovirus, the only other nonenveloped virus studied by this method, shows the optimum rate of marker release from liposomes at around pH 6 (R. Blumenthal, P. S. Seth, M. C. Willingham, and I. Pastan, Biochemistry 25:2231-2237, 1986). The interaction between rotavirus and liposomes is sensitive to specific divalent metal ions, unlike the adenovirus-liposome interaction, which is independent of them.
Nandi
P
P
Unité de Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, Nouzilly, France.
Charpilienne
A
A
Cohen
J
J
D000818:Animals; D000956:Antigens, Viral; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D002413:Cations, Divalent / Q000494:pharmacology; D002417:Cattle / Q000382:microbiology; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism*; D008954:Models, Biological; D012401:Rotavirus / Q000187:drug effects / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology; D015678:Viral Structural Proteins / Q000378:metabolism*
D000956:Antigens, Viral; D036022:Capsid Proteins; D002413:Cations, Divalent; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008081:Liposomes; C060339:VP4 protein, Rotavirus; C050779:VP7 protein, Rotavirus; D015678:Viral Structural Proteins; C024098:6-carboxyfluorescein; D014357:Trypsin