EEF1A binding to aminoacylated viral RNA represses minus strand synthesis by TYMV RNA-dependent RNA polymerase

Abstract

The genomic RNA of Turnip yellow mosaic virus (TYMV) has an 82-nucleotide-long tRNA-like structure at its 3′-end that can be valylated and then form a stable complex with translation elongation factor eEF1A·GTP. Transcription of this RNA by TYMV RNA-dependent RNA polymerase (RdRp) to yield minus strands has previously been shown to initiate within the 3′-CCA sequence. We have now demonstrated that minus strand synthesis is strongly repressed upon the binding of eEF1A·GTP to the valylated viral RNA. eEF1A·GTP had no effect on RNA synthesis templated by non-aminoacylated RNA. Higher eEF1A·GTP levels were needed to repress minus strand synthesis templated by valyl-EMV TLS RNA, which binds eEF1A·GTP with lower affinity than does valyl-TYMV RNA. Repression by eEF1A·GTP was also observed with a methionylated variant of TYMV RNA and with aminoacylated tRNA His, tRNA Ala, and tRNA Phe transcripts. It is proposed that minus strand repression by eEF1A·GTP binding occurs early during infection to help coordinate the competing translation and replication functions of the genomic RNA.