Effect of expression of the aphthovirus protease 3C on viral infection and gene expression.
Cells transformed with specific regions of the foot-and-mouth disease virus (FMDV) genome have been constructed and analyzed with respect to viability and susceptibility to FMDV infection. Constitutive expression of an active protease 3C under the control of the tk promoter has been documented by the ability of transformed cells to catalyze the processing of a P1 capsid precursor. High-level, transient expression but not low-level, constitutive expression, of 3C caused a 10-fold reduction in the yield of FMDV and was detrimental to the expression of the cotransfected reporter luciferase gene. No such effect was observed in assays involving cells transfected with a deleted, inactive form of 3C. The negative effect of 3C was not observed when the same reporter gene was integrated and expressed in a constitutive fashion nor when its translation was directed by the internal ribosome entry site element of FMDV in transient expression assays. The results show that cells with a low level of expression of the aphthoviral 3C can be stably maintained and can provide a useful tool to study polyprotein processing.
Martínez-Salas
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Centro de Biología Molecular Severo Ochoa (CSIC, UAM), Universidad Autónoma de Madrid, Spain.
Domingo
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D000086042:3C Viral Proteases; D000595:Amino Acid Sequence; D000818:Animals; D005537:Aphthovirus / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D015967:Gene Expression Regulation, Viral; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010442:Peptide Chain Initiation, Translational; D012333:RNA, Messenger / Q000235:genetics; D014764:Viral Proteins; D014779:Virus Replication
D017931:DNA Primers; D012333:RNA, Messenger; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases