Stoichiometry of murine leukemia virus envelope protein-mediated fusion and its neutralization.
Envelope glycoproteins (Envs) of retroviruses form trimers that mediate fusion between viral and cellular membranes and are the targets for neutralizing antibodies. Understanding in detail how Env trimers mediate membrane fusion, and how antibodies interfere with this process, is a fundamental problem in biology with practical implications for the development of antiviral drugs and vaccines. We investigated the stoichiometry of Env-mediated fusion and its inhibition by antibody by inserting an epitope from human immunodeficiency virus for a neutralizing antibody (2F5) into the surface (SU) or transmembrane (TM) protein of murine leukemia virus Env, along with point mutations that abrogate SU and TM function but complement one another. We transfected various combinations of these Env genes and investigated Env-mediated cell fusion and its inhibition by 2F5 antibody. Our results showed that heterotrimers with one functional SU molecule were fusion competent in complementation experiments and that one antibody molecule was sufficient to inactivate the fusion function of a trimer when its epitope was in functional SU or TM. 2F5 antibody could also neutralize trimers with the 2F5 epitope in nonfunctional SU or TM, but less efficiently.
Ou
Wu
W
Building 4/Room 336, 4 Center Dr., Bethesda, MD 20892, USA.
Silver
Jonathan
J
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D005816:Genetic Complementation Test; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics*; D016296:Mutagenesis; D009500:Neutralization Tests; D014760:Viral Fusion Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D053586:Virus Internalization
D000911:Antibodies, Monoclonal; D014760:Viral Fusion Proteins