Truncation of the COOH-terminal region of the paramyxovirus SV5 fusion protein leads to hemifusion but not complete fusion.
The role of the simian virus 5 (SV5) fusion (F) protein 20 residue COOH-terminal region, thought to represent the cytoplasmic tail, in fusion activity was examined by constructing a series of COOH-terminal truncation mutants. When the altered F proteins were expressed in eukaryotic cells, by using the vaccinia virus-T7 transient expression system, all the F proteins exhibited similar intracellular transport properties and all were expressed abundantly on the cell surface. Quantitative and qualitative cell fusion assays indicated that all of the F protein COOH-terminal truncation mutants mediated lipid mixing with similar kinetics and efficiency as that of wild-type F protein. However, the cytoplasmic content mixing activity decreased in parallel with the extent of the deletion in the F protein COOH-terminal truncation mutants. These data indicate that it is possible to separate the presumptive early step in the fusion reaction, hemifusion, and the final stage of fusion, content mixing, and that the presence of the F protein COOH-terminal region is important for the final steps of fusion.
Bagai
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S
Howard Hughes Medical Institute, Northwestern University, Evanston, Illinois 60208-3500, USA.
Lamb
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RA
D000595:Amino Acid Sequence; D001692:Biological Transport; D015496:CD4-Positive T-Lymphocytes; D050260:Carbohydrate Metabolism; D002459:Cell Fusion; D002462:Cell Membrane / Q000737:chemistry / Q000502:physiology; D003593:Cytoplasm; D019281:Dimerization; D004910:Erythrocyte Membrane / Q000502:physiology; D005456:Fluorescent Dyes; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006596:Hexosaminidases; D006801:Humans; D007700:Kinetics; D008561:Membrane Fusion / Q000502:physiology*; D008563:Membrane Lipids / Q000502:physiology; D008969:Molecular Sequence Data; D010252:Paramyxoviridae / Q000502:physiology*; D017384:Sequence Deletion; D014760:Viral Fusion Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
D005456:Fluorescent Dyes; D008563:Membrane Lipids; D014760:Viral Fusion Proteins; D006596:Hexosaminidases