title: The fusion-controlling disulfide bond isomerase in retrovirus Env is triggered by protein destabilization.
abstract: The membrane fusion function of murine leukemia virus (MLV) is carried by the Env protein. This protein is composed of three SU-TM subunit complexes. The fusion activity is loaded into the transmembrane TM subunit and controlled by the peripheral, receptor-binding SU subunit. It is assumed that TM adopts a metastable conformation in the native Env and that fusion activation involves the folding of TM into a stable form. Activation is suppressed by the associated SU and triggered by its dissociation, which follows receptor binding. Recently we showed that the two subunits are disulfide linked and that SU dissociation and triggering of the fusion function are caused by a switch of the intersubunit disulfide into an intrasubunit disulfide isomer using an isomerization-active CWLC motif in SU (M. Wallin, M. Ekstrom, and H. Garoff, EMBO J. 23:54-65, 2004). In the present work we address how the SU disulfide isomerase is activated. Using Moloney MLV, we show that isomerization of the SU-TM disulfide bond can be triggered by heat, urea, or guanidinium hydrochloride. Such protein perturbation treatments also significantly increase the kinetics and efficiency of viral fusion. The threshold conditions for the effects on isomerization and fusion are virtually the same. This finding indicates that destabilization of interactions in the SU oligomer induces the disulfide bond isomerase and the subsequent activation of the fusion function in TM.
authors.affiliation: Department of Biosciences at Novum, Karolinska Institute, S-141 57 Huddinge, Sweden.
authors.forename: Michael
authors.initials: M
authors.lastname: Wallin
authors.forename: Maria
authors.initials: M
authors.lastname: Ekström
authors.forename: Henrik
authors.initials: H
authors.lastname: Garoff
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004220:Disulfides / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D015686:Gene Products, env / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019791:Guanidine / Q000494:pharmacology; D006358:Hot Temperature; D008561:Membrane Fusion / Q000502:physiology*; D051379:Mice; D008979:Moloney murine leukemia virus / Q000187:drug effects / Q000378:metabolism / Q000472:pathogenicity*; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D013237:Stereoisomerism; D014508:Urea / Q000494:pharmacology
chemical_list: D004220:Disulfides; D015686:Gene Products, env; D014508:Urea; D019704:Protein Disulfide-Isomerases; D019791:Guanidine