Sequence motif upstream of the Hendra virus fusion protein cleavage site is not sufficient to promote efficient proteolytic processing.
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.
Craft
Willie Warren
WW
Department of Molecular and Cellular Biochemistry, University of Kentucky, 741 South Limestone, Biomedical Biological Sciences Research Building, Lexington, KY 40536-0509, USA.
Dutch
Rebecca Ellis
RE
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D045404:Hendra Virus / Q000235:genetics* / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014357:Trypsin / Q000378:metabolism / Q000494:pharmacology; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins; D014357:Trypsin