Site-directed mutagenesis of Avibirnavirus VP4 gene.
Virus protein VP4 of infectious bursal disease virus (IBDV) is a protease which separates VPX and VP3 from the polyprotein. We studied the importance of serine and aspartic acid on cleavage at the VPX/VP4 junction and analysed the role of the proposed H547, D590, and S653 catalytic site using five different mutations on VP4. Our results suggest that the replacement of serine by lysine in AXAAS motifs in serotype II IBDV influences polyprotein (PP) processing by VP4 and also indicate the presence of an alternative cleavage site. Mutation on D ((510)TLAADK(515)) prevented the cleavage at the VPX/VP4 junction, but we have found that independently of the importance of those alanines in LAA, D has an important role as part of the cleavage site. Replacement of histidine by proline H547P completely abolished PP processing. Mutation on D590 induced a partial PP processing when it was replaced by proline and the replacement of serine by proline at S653P induced a prominent change in PP processing. These results permit us to conclude that IBDV VP4 has the ability to act according to structural and topographical changes during translational and posttranslational processes and allow multiple hit sites, which serve to increase effectiveness.
Rodríguez-Lecompte
Juan Carlos
JC
Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island, Canada C1A 4P3.
Kibenge
Frederick S B
FS
D000595:Amino Acid Sequence; D007243:Infectious bursal disease virus / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D014176:Protein Biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics*; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic
D015534:Trans-Activators; D012697:Serine Endopeptidases; C404159:virus protein 4, infectious bursal disease virus strain P2