Variable sensitivity to substitutions in the N-terminal heptad repeat of Mason-Pfizer monkey virus transmembrane protein.
The transmembrane protein of Mason-Pfizer monkey virus contains two heptad repeats that are predicted to form amphipathic alpha-helices that mediate the conformational change necessary for membrane fusion. To analyze the relative sensitivity of the predicted hydrophobic face of the N-terminal heptad repeat to the insertion of uncharged, polar, and charged substitutions, mutations that introduced alanine, serine, or glutamic acid into positions 436, 443, 450, and 457 of the envelope protein were examined. Novel systems using Tat protein and the GHOST cell line were developed to test and quantitate the effects of the mutations on Env-mediated fusion and infectivity of the virus. While no single amino acid change at any of the positions interfered significantly with the synthesis, processing, or transport to the plasma membrane of glycoprotein complexes, 9 of the 12 nonconservative mutations in these residues completely abolished fusion activity and virus infectivity. Mutations in the central positions (443 and 450) of the heptad repeat region were the most detrimental to Env function, and even single alanine substitutions in these positions dramatically altered the fusogenicity of the protein. These results demonstrate that this N-terminal heptad repeat plays a critical role in Env-mediated membrane fusion and highlight the key function of central hydrophobic residues in this process and the sensitivity of all positions to charge substitutions.
Song
Chisu
C
Department of Microbiology and Center for AIDS Research, University of Alabama at Birmingham, 845 19th Street S., Birmingham, AL 35294, USA.
Hunter
Eric
E
D020816:Amino Acid Motifs; D019943:Amino Acid Substitution; D000818:Animals; D019556:COS Cells; D002459:Cell Fusion; D016093:Mason-Pfizer monkey virus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D008561:Membrane Fusion; D016297:Mutagenesis, Site-Directed; D020449:Repetitive Sequences, Amino Acid; D020079:Terminal Repeat Sequences; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism
D014759:Viral Envelope Proteins