Virus-mediated release of endosomal content in vitro: different behavior of adenovirus and rhinovirus serotype 2.
Endosomal penetration by nonenveloped viruses might be accomplished by either local breakdown of the endosomal membrane (e.g., adenovirus) or formation of a membrane-spanning pore by capsid proteins. Uncoating of the nonenveloped virus human rhinovirus serotype 2 (HRV2) has been shown to occur from late endosomes and to be entirely dependent on the acidic pH in this compartment (Prchla, E., E. Kuechler, D. Blaas, and R. Fuchs. 1994. J. Virol. 68: 3713-3723). To investigate further the mechanism of uncoating of HRV2, an in vitro assay was established to test viruses or virus-derived peptides for their capacity to release cointernalized biotin-dextran of different molecular mass (10 and 70 kD) from isolated endosomes. The suitability of the assay was demonstrated by use of a fusogenic peptide derived from influenza virus hemagglutinin (GALA-INF3). Whereas adenovirus induced a low pH-dependent release of up to 46% of the internalized biotin-dextran and did not show any significant size selectivity (as expected for endosome disruption), HRV2 mediated release of 27% of the 10 kD dextran and only traces of the 70-kD dextran. Similarly, GALA-INF3-induced release of biotin-dextran was also size dependent. The potential role of the capsid protein VP1 in HRV2 uncoating in vivo was also substantiated in our in vitro system using an amphipathic, NH2-terminal peptide of VP1. Taken together, these data favor the model of a specific pore-forming mechanism for HRV2 uncoating which is in contrast to the membrane-disrupting mechanism of adenovirus.
Prchla
E
E
Department of General and Experimental Pathology, University of Vienna, Austria.
Plank
C
C
Wagner
E
E
Blaas
D
D
Fuchs
R
R
D000256:Adenoviridae / Q000378:metabolism*; D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000378:metabolism; D015415:Biomarkers; D001710:Biotin; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins; D002474:Cell-Free System / Q000378:metabolism / Q000821:virology; D003911:Dextrans; D011992:Endosomes / Q000378:metabolism* / Q000821:virology*; D006367:HeLa Cells / Q000378:metabolism / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000494:pharmacology; D018272:Porins / Q000378:metabolism; D012229:Rhinovirus / Q000145:classification / Q000378:metabolism*; D012703:Serotyping
D000956:Antigens, Viral; D015415:Biomarkers; D036022:Capsid Proteins; D003911:Dextrans; D010455:Peptides; D018272:Porins; D001710:Biotin