Sequence motif upstream of the Hendra virus fusion protein cleavage site is not sufficient to promote efficient proteolytic processing

• Published in: Virology (New York, N.Y.) N.Y.), 2005-10, Vol.341 (1), p.130-140
• Main Authors: Craft, Willie Warren, Dutch, Rebecca Ellis
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; ALANINES; Amino Acid Motifs; Amino Acid Sequence; Animals; Binding Sites - genetics; Cell Line; Cercopithecus aethiops; CLEAVAGE; Cricetinae; Fusion protein; Hendra; Hendra virus; Hendra Virus - genetics; Hendra Virus - metabolism; Henipavirus; horses; MODIFICATIONS; Morbillivirus; Mutagenesis, Site-Directed; MUTANTS; Paramyxovirus; post-translational modification; Post-translational modifications; PROTEIN ENGINEERING; Protein Processing, Post-Translational; PROTEINS; proteolysis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Transfection; Trypsin - metabolism; Trypsin - pharmacology; Vero Cells; viral fusion proteins; Viral Fusion Proteins - chemistry; Viral Fusion Proteins - genetics; Viral Fusion Proteins - metabolism; VIRUSES
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/j.virol.2005.07.004

The Hendra virus fusion (HeV F) protein is synthesized as a precursor, F 0, and proteolytically cleaved into the mature F 1 and F 2 heterodimer, following an HDLVDGVK 109 motif. This cleavage event is required for fusogenic activity. To determine the amino acid requirements for processing of the HeV F protein, we constructed multiple mutants. Individual and simultaneous alanine substitutions of the eight residues immediately upstream of the cleavage site did not eliminate processing. A chimeric SV5 F protein in which the furin site was substituted for the VDGVK 109 motif of the HeV F protein was not processed but was expressed on the cell surface. Another chimeric SV5 F protein containing the HDLVDGVK 109 motif of the HeV F protein underwent partial cleavage. These data indicate that the upstream region can play a role in protease recognition, but is neither absolutely required nor sufficient for efficient processing of the HeV F protein.