Physiologic RNA Targets and Refined Sequence Specificity of Coronavirus EndoU

BioRxiv : the Preprint Server for Biology
R. AncarDavid L Barton


Coronavirus EndoU inhibits dsRNA-activated antiviral responses; however, the physiologic RNA substrates of EndoU are unknown. In this study, we used mouse hepatitis virus (MHV)-infected bone-marrow-derived macrophage (BMM) and cyclic phosphate cDNA sequencing to identify the RNA targets of EndoU. EndoU targeted viral RNA, cleaving the 3'-side of pyrimidines with a strong preference for U|A and C|A sequences (endoY|A). EndoU-dependent cleavage was detected in every region of MHV RNA, from the 5'-NTR to the 3'-NTR, including transcriptional regulatory sequences (TRS). Cleavage at two CA dinucleotides immediately adjacent to the MHV poly(A) tail suggest a mechanism to suppress negative-strand RNA synthesis and the accumulation of viral dsRNA. MHV with EndoU or 2-5A phosphodiesterase mutations provoked the activation of RNase L in BMM, with corresponding cleavage of RNAs by RNase L. The physiologic targets of EndoU are viral RNA templates required for negative-strand RNA synthesis and dsRNA accumulation.

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