Theoretical and functional analysis of the SIV fusion peptide.
The fusion domain of simian immunodeficiency virus (SIV) envelope glycoproteins is a hydrophobic region located at the amino-terminal extremity of the transmembrane protein (gp32). Assuming an alpha helical structure for the SIV fusogenic domain of gp32 in a lipid environment, theoretical studies have predicted that the fusion peptide would insert obliquely in the lipid bilayer. This oblique insertion could be an initial step of the fusion process by disorganizing locally the structure of the lipid bilayer. We have tested this hypothesis by selectively mutagenizing the SIV gp160 expressed via a vaccinia virus vector, to alter the theoretical angle of insertion of the fusion peptide. The fusogenic activity of the wild-type and mutant glycoproteins was tested after infection of T4 lymphocytic cell lines by the recombinant vaccinia virus, and measure of syncytia formation. Mutations that modified the oblique orientation reduced the fusogenic activity. In contrast, mutations that conserve the oblique orientation did not alter the fusogenic properties. Our results support the hypothesis that oblique orientation is important for fusogenic activity.
Horth
M
M
Laboratory of Biological Chemistry, Free University of Brussels, Belgium.
Lambrecht
B
B
Khim
M C
MC
Bex
F
F
Thiriart
C
C
Ruysschaert
J M
JM
Burny
A
A
Brasseur
R
R
D000595:Amino Acid Sequence; D000818:Animals; D015704:CD4 Antigens / Q000378:metabolism; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011233:Precipitin Tests; D011487:Protein Conformation; D015531:Radioimmunoprecipitation Assay; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism*
D015704:CD4 Antigens; D006023:Glycoproteins; D008051:Lipid Bilayers; D014760:Viral Fusion Proteins