Proteolytic activation of respiratory syncytial virus fusion protein. Cleavage at two furin consensus sequences.
The F (fusion) protein of the respiratory syncytial viruses is synthesized as an inactive precursor F(0) that is proteolytically processed at the multibasic sequence KKRKRR(136) into the subunits F(1) and F(2) by the cellular protease furin. This maturation process is essential for the F protein to gain fusion competence. We observed that proteolytic cleavage additionally occurs at another basic motif, RARR(109), that also meets the requirements for furin recognition. Cleavage at both sites leads to the removal from the polypeptide chain of a glycosylated peptide of 27 amino acids. When the sequence RARR(109) was changed to NANR(109) or to RANN(109) by site-directed mutagenesis, cleavage by furin was completely prevented. Although the mutants were still processed at position Arg(136), they did not show any syncytia formation. Proteolytic cleavage of the modified motifs was achieved by treatment of transfected cells with trypsin converting the F mutants into their fusogenic forms. Our findings indicate that both furin consensus sequences have to be cleaved in order to activate the fusion protein.
Zimmer
G
G
Institut für Virologie, Tierärztliche Hochschule Hannover, Bünteweg 17, D-30559 Hannover, Germany.
Budz
L
L
Herrler
G
G
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D045683:Furin; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D012136:Respiratory Syncytial Viruses / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D013381:Subtilisins / Q000378:metabolism*; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry*
D014760:Viral Fusion Proteins; D014764:Viral Proteins; D013381:Subtilisins; D045683:Furin