Mutations in a carboxy-terminal region of vesicular stomatitis virus glycoprotein G that affect membrane fusion activity.
The envelope glycoprotein G of vesicular stomatitis virus induces membrane fusion at acidic pH. A highly conserved amino terminal region spanning residues 123 to 137 has previously been identified as an internal fusion domain. Here we have substituted specific amino acids within a carboxy terminal region, conserved in five vesiculoviruses encompassing residues 395 to 418, and studied the effect of these mutations on membrane fusion at acid pH and pH-dependent conformational change. Substitution of conserved Gly 395, Gly 404, Gly 406, Asp 409, and Asp 411 with Glu, Ala, Ala, Asn, and Asn, respectively, decreased the cell-cell fusion efficiency, as well as reduced the pH threshold of membrane fusion. Mutation of Gly 404 and Asp 409 to Lys and Ala, respectively, abolished the fusion activity. Mutant Gly 404 Lys also showed markedly altered resistance to trypsin digestion at acidic pH. These results suggest that the region between amino acids 395 to 418 is important for the fusogenic activity of the G protein. The possible role of this domain in conformational changes involved in fusion activity of VSV G is also discussed.
Shokralla
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Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
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Wanas
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Ghosh
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D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002459:Cell Fusion / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology; D003001:Cloning, Molecular; D017124:Conserved Sequence; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008561:Membrane Fusion; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D014357:Trypsin; D014721:Vesicular stomatitis Indiana virus / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014357:Trypsin