Dimerization of glycoprotein E(rns) of classical swine fever virus is not essential for viral replication and infection.
The pestivirus glycoprotein E(rns), a ribonuclease, is expressed on the surface of virions and in infected cells as a disulfide-linked homodimer. E(rns) is involved in the infection process and its RNase activity is probably involved in viral replication and pathogenesis. The most C-terminal cysteine residue forms an intermolecular disulfide bond with another E(rns) monomer, resulting in an E(rns) dimer. To study the function of dimerisation of E(rns) for viral replication, the cysteine residue at amino acid position 438 was mutated into a serine residue. The mutated C438S gene was cloned into a vector containing an infectious cDNA copy of the CSFV C-strain genome. Using reverse genetics, a mutant virus was generated that only expressed monomeric E(rns), confirming that Cys 438 is essential for homo-dimerization. Characterization of this mutant virus and of a baculovirus-expressed C438S mutant protein indicated that the loss of the dimeric state of E(rns) reduced the affinity of binding of virions and E(rns) to heparan sulphate (HS), the receptor for E(rns) on the cell surface of SK6 cells. This suggests that interaction of virus-bound E(rns) homodimers with membrane associated HS may be a joined action of the two HS-binding domains (one in each monomer) present in the homodimer.
Animal Sciences Group (Wageningen UR, former ID-Lelystad), Lelystad, The Netherlands. RENE.VANGENNIP@WUR.NL
H G P
HG
van Gennip
A T
AT
Hesselink
R J M
RJ
Moormann
M M
MM
Hulst
D000818:Animals; D002460:Cell Line; D006691:Classical Swine Fever / Q000821:virology*; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D017931:DNA Primers; D019281:Dimerization; D007668:Kidney; D008969:Molecular Sequence Data; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism; D014779:Virus Replication / Q000502:physiology*
D017931:DNA Primers; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C089580:glycoprotein E, Suid herpesvirus 1