VPg-primed RNA synthesis of norovirus RNA-dependent RNA polymerases by using a novel cell-based assay.

ABSTRACT: Molecular studies of human noroviruses (NoV) have been hampered by the lack of a permissive cell culture system. We have developed a sensitive and reliable mammalian cell-based assay for the human NoV GII.4 strain RNA-dependent RNA polymerase (RdRp). The assay is based on the finding that RNAs synthesized by transiently expressed RdRp can stimulate retinoic acid-inducible gene I (RIG-I)-dependent reporter luciferase production via the beta interferon promoter. Comparable activities were observed for the murine norovirus (MNV) RdRp. RdRps with mutations at divalent metal ion binding residues did not activate RIG-I signaling. Furthermore, both NoV and MNV RdRp activities were stimulated by the coexpression of their respective VPg proteins, while mutations in the putative site of nucleotide l

SUBMITTER: Subba-Reddy CV 

PROVIDER: S-EPMC3233154 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature