Structural and functional analysis of the HIV gp41 core containing an Ile573 to Thr substitution: implications for membrane fusion.
The envelope glycoprotein of HIV-1 consists of the surface subunit gp120 and the transmembrane subunit gp41. Binding of gp120 to target cell receptors induces a conformational change in gp41, which then mediates the fusion of viral and cellular membranes. A buried isoleucine (Ile573) in a central trimeric coiled coil within the fusion-active gp41 ectodomain core is thought to favor this conformational activation. The role of Ile573 in determining the structure and function of the gp120-gp41 complex was investigated by mutating this residue to threonine, a nonconservative substitution in HIV-1 that occurs naturally in SIV. While the introduction of Thr573 markedly destabilized the gp41 core, the three-dimensional structure of the mutant trimer of hairpins was very similar to that of the wild-type molecule. A new hydrogen-bonding interaction between the buried Thr573 and Thr569 residues appears to allow formation of the trimer-of-hairpins structure at physiological temperature. The mutant envelope glycoprotein expressed in 293T cells and incorporated within pseudotyped virions displayed only a moderate reduction in syncytium-inducing capacity and virus infectivity, respectively. Our results demonstrate that the proper folding of the gp41 core underlies the membrane fusion properties of the gp120-gp41 complex. An understanding of the gp41 activation process may suggest novel strategies for vaccine and antiviral drug development.
Department of Biochemistry, Weill Medical College of Cornell University, New York, New York 10021, USA.
J
J
Liu
W
W
Shu
M B
MB
Fagan
J H
JH
Nunberg
M
M
Lu
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics*; D002460:Cell Line; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D015497:HIV-1 / Q000235:genetics* / Q000472:pathogenicity; D006801:Humans; D007532:Isoleucine / Q000235:genetics*; D008561:Membrane Fusion / Q000235:genetics*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D020449:Repetitive Sequences, Amino Acid; D013329:Structure-Activity Relationship; D013912:Threonine / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014771:Virion / Q000235:genetics / Q000472:pathogenicity; D014774:Virulence
D015700:HIV Envelope Protein gp41; D007532:Isoleucine; D013912:Threonine