Deletion mutants of VPg reveal new cytopathology determinants in a picornavirus.
BACKGROUND
Success of a viral infection requires that each infected cell delivers a sufficient number of infectious particles to allow new rounds of infection. In picornaviruses, viral replication is initiated by the viral polymerase and a viral-coded protein, termed VPg, that primes RNA synthesis. Foot-and-mouth disease virus (FMDV) is exceptional among picornaviruses in that its genome encodes 3 copies of VPg. Why FMDV encodes three VPgs is unknown.


RESULTS
we have constructed four mutant FMDVS that encode only one VPG: either VPg(1), VPg(3), or two chimeric versions containing part of VPg(1) and VPg(3). All mutants, except that encoding only VPg(1), were replication-competent. Unexpectedly, despite being replication-competent, the mutants did not form plaques on BHK-21 cell monolayers. The one-VPg mutant FMDVs released lower amounts of encapsidated viral RNA to the extracellular environment than wild type FMDV, suggesting that deficient plaque formation was associated with insufficient release of infectious progeny. Mutant FMDVs subjected to serial passages in BHK-21 cells regained plaque-forming capacity without modification of the number of copies of VPg. Substitutions in non-structural proteins 2C, 3A and VPg were associated with restoration of plaque formation. Specifically, replacement R55W in 2C was repeatedly found in several mutant viruses that had regained competence in plaque development. The effect of R55W in 2C was to mediate an increase in the extracellular viral RNA release without a detectable increase of total viral RNA that correlated with an enhanced capacity to alter and detach BHK-21 cells from the monolayer, the first stage of cell killing.


CONCLUSIONS
The results link the VPg copies in the FMDV genome with the cytopathology capacity of the virus, and have unveiled yet another function of 2C: modulation of picornavirus cell-to-cell transmission. Implications for picornaviruses pathogenesis are discussed.
Arias
Armando
A
Departamento de Virología y Microbiología, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Consejo Superior de Investigaciones Científicas (CSIC), Campus de Cantoblanco, Madrid, Spain.
Perales
Celia
C
Escarmís
Cristina
C
Domingo
Esteban
E
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral / Q000235:genetics*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D008969:Molecular Sequence Data; D010641:Phenotype; D010849:Picornaviridae / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D012367:RNA, Viral / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
D012367:RNA, Viral; D014764:Viral Proteins