Conformational alteration of Sindbis virion glycoproteins induced by heat, reducing agents, or low pH.
Sindbis virions undergo a conformational rearrangement after attachment to cells but prior to entry, as detected by exposure of epitopes on virus-cell complexes which are not accessible to their cognate monoclonal antibodies on native virions (D. C. Flynn, W. J. Meyer, and R. E. Johnston, J. Virol. 64:3643-3653, 1990). The rearrangement did not appear to require transit of virions through a low-pH environment, and the altered virions participated in a productive infection. This naturally occurring structural alteration could be mimicked, although not precisely duplicated, by any of the three artificial treatments of purified virions in vitro: brief incubation at 51 degrees C, treatment with 1 to 5 mM dithiothreitol, or incubation of pH 5.8 to 6.0. Infectivity was maintained after all three treatments, suggesting that Sindbis virions are metastable and can exist in at least two infectious conformations. The integrity of external, neutralizing epitopes was maintained on cell-associated virions and in the altered conformations induced by heat and dithiothreitol, whereas these epitopes were unreactive under low-pH conditions that induced an analogous exposure of previously inaccessible epitopes. The pH at which the conformational change was induced and the pH at which virions could mediate cell-cell fusion from without were coordinately shifted when these two parameters were determined for another strain of Sindbis virus. This coordinate shift in pH optima suggests that the conformational change in virion structure observed at the cell surface may be causally related to fusion.
Meyer
W J
WJ
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill 27599-7290.
Gidwitz
S
S
Ayers
V K
VK
Schoepp
R J
RJ
Johnston
R E
RE
D000911:Antibodies, Monoclonal; D002459:Cell Fusion; D004229:Dithiothreitol / Q000494:pharmacology; D000939:Epitopes / Q000032:analysis; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation; D011489:Protein Denaturation; D012845:Sindbis Virus / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure*; D014759:Viral Envelope Proteins / Q000187:drug effects / Q000276:immunology*; D014771:Virion / Q000737:chemistry / Q000472:pathogenicity / Q000648:ultrastructure*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D014759:Viral Envelope Proteins; C064950:glycoprotein E2, Sindbis virus; D004229:Dithiothreitol