Mechanism of adenovirus-mediated endosome lysis: role of the intact adenovirus capsid structure.
Adenoviruses have been previously shown to enhance the delivery of many ligands including proteins and plasmid DNAs to the cells. The key biochemical step during this process is the ability of adenovirus to disrupt (lyse) the endosome membrane releasing the co-internalized virus and the other ligands into the cytosol (Seth et al, 1986, In: Adenovirus attachment and entry into cells, pp 191-195, American Society for Microbiology, Washington, D.C.). To understand the role of the adenovirus proteins involved in the endosome lysis, it is further shown here that empty capsids of adenovirus also possess this membrane vesicle lytic activity; though the activity is about 5-times lower than the adenovirus. Incubation of adenovirus with low concentration of ionic detergent or brief exposure to 45 degrees C destroyed this lytic activity without affecting the adenovirus binding to cell surface receptor, suggesting the lytic activity of adenovirus to be of enzymatic nature. However, exposing adenovirus to conditions that can disrupt adenovirus capsid structure such as heating at 65 degrees C, treating with 0.5% SDS, treating with different proteases, dialyzing against no glycerol buffer, treating with 6 M urea or with 10% pyridine, and sonication destroyed the adenovirus-associated lytic activity. Results suggest the requirement of an intact capsid structure for adenovirus-mediated lysis of the endosome.
Seth
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Laboratory of Molecular Biology, NCI, NIH, Bethesda, MD 20892.
D000260:Adenoviruses, Human / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D002213:Capsid / Q000302:isolation & purification / Q000378:metabolism*; D003600:Cytosol; D003956:Dialysis; D010450:Endopeptidases / Q000494:pharmacology; D011992:Endosomes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007624:KB Cells; D008715:Methionine / Q000378:metabolism; D013462:Sulfur Radioisotopes; D014465:Ultrasonics
D013462:Sulfur Radioisotopes; D008715:Methionine; D010450:Endopeptidases