Diphosphates at the 5′ end of the positive strand of yeast L-A double-stranded RNA virus as a molecular self-identity tag

Abstract

The 5′end of RNA conveys important information on self-identity. In mammalian cells, double-stranded RNA (dsRNA) with 5′di- or triphosphates generated during virus infection is recognized as foreign and elicits the host innate immune response. Here, we analyze the 5′ ends of the dsRNA genome of the yeast L-A virus. The positive strand has largely diphosphates with a minor amount of triphosphates, while the negative strand has only diphosphates. Although the virus can produce capped transcripts by cap snatching, neither strand carried a cap structure, suggesting that only non-capped transcripts serve as genomic RNA for encapsidation. We also found that the 5′ diphosphates of the positive but not the negative strand within the dsRNA genome are crucial for transcription in vitro. Furthermore, the presence of a cap structure in the dsRNA abrogated its template activity. Given that the 5′ diphosphates of the transcripts are also essential for cap acquisition and that host cytosolic RNAs (mRNA, rRNA, and tRNA) are uniformly devoid of 5′ pp-structures, the L-A virus takes advantage of its 5′ terminal diphosphates, using them as a self-identity tag to propagate in the host cytoplasm.