A single amino acid change in the Newcastle disease virus fusion protein alters the requirement for HN protein in fusion.
The role of a leucine heptad repeat motif between amino acids 268 and 289 in the structure and function of the Newcastle disease virus (NDV) F protein was explored by introducing single point mutations into the F gene cDNA. The mutations affected either folding of the protein or the fusion activity of the protein. Two mutations, L275A and L282A, likely interfered with folding of the molecule since these proteins were not proteolytically cleaved, were minimally expressed at the cell surface, and formed aggregates. L268A mutant protein was cleaved and expressed at the cell surface although the protein migrated slightly slower than wild type on polyacrylamide gels, suggesting an alteration in conformation or processing. L268A protein was fusion inactive in the presence or absence of HN protein expression. Mutant L289A protein was expressed at the cell surface and proteolytically cleaved at better than wild-type levels. Most importantly, this protein mediated syncytium formation in the absence of HN protein expression although HN protein enhanced fusion activity. These results show that a single amino acid change in the F(1) portion of the NDV F protein can alter the stringent requirement for HN protein expression in syncytium formation.
Sergel
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Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
McGinnes
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LW
Morrison
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TG
D000409:Alanine / Q000235:genetics; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D019556:COS Cells; D015707:HN Protein / Q000378:metabolism; D007930:Leucine / Q000235:genetics / Q000502:physiology*; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009522:Newcastle disease virus / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011499:Protein Processing, Post-Translational; D014760:Viral Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D015707:HN Protein; D014760:Viral Fusion Proteins; D007930:Leucine; D000409:Alanine