Transient inhibition of foot-and-mouth disease virus infection of BHK-21 cells by antisense oligonucleotides directed against the second functional initiator AUG.
The antiviral activity of antisense oligonucleotides corresponding to different regions of foot-and-mouth disease virus (FMDV) genome has been assessed in BHK-21 cells. The locations of the oligonucleotides used were: (i) two regions within the internal ribosome entry site (IRES), involved in the regulation of the translation initiation of the viral polyprotein; (ii) each of the two functional initiator AUGs; (iii) an internal sequence of P2A gene; and (iv) a region at the 3' end non-coding region. Cytoplasmic microinjection of oligodeoxyribonucleotides and oligoribonucleotides complementary to the second AUG resulted in a transient inhibition of viral VP1 expression in infected cells. Significant inhibitions, ranging from 35 to 52%, were obtained at 5 h post-infection using oligonucleotide concentrations of 125 microM and higher. The extent and duration of this inhibition seemed to be mediated by both a rapid transport to the nucleus and the short half-life of the oligonucleotide. This inhibition of FMDV protein synthesis was correlated with a reduction of virus yield of about 50%, as observed after the addition to the cell culture of an oligodeoxyribonucleotide phosphorothioate complementary to the second AUG.
Gutiérrez
A
A
Departamento de Producción Animal, CIT-INIA, Madrid, Spain.
Rodríguez
A
A
Pintado
B
B
Sobrino
F
F
D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use*; D005537:Aphthovirus / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line / Q000382:microbiology; D003062:Codon / Q000187:drug effects / Q000235:genetics; D003470:Culture Media; D005536:Foot-and-Mouth Disease / Q000517:prevention & control*; D008845:Microinjections; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000627:therapeutic use*; D010448:Peptide Initiation Factors / Q000187:drug effects / Q000235:genetics*; D014176:Protein Biosynthesis / Q000187:drug effects / Q000235:genetics; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D003062:Codon; D003470:Culture Media; D016376:Oligonucleotides, Antisense; D010448:Peptide Initiation Factors; D014764:Viral Proteins