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The predicted stem-loop structure in the 3'-end of the human norovirus antigenomic sequence is required for its genomic RNA synthesis by its RdRp.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2021 Oct; Vol. 297 (4), pp. 101225. Date of Electronic Publication: 2021 Sep 23. - Publication Year :
- 2021
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Abstract
- The norovirus genome consists of a single positive-stranded RNA. The mechanism by which this single-stranded RNA genome is replicated is not well understood. To reveal the mechanism underlying the initiation of the norovirus genomic RNA synthesis by its RNA-dependent RNA polymerase (RdRp), we used an in vitro assay to detect the complementary RNA synthesis activity. Results showed that the purified recombinant RdRp was able to synthesize the complementary positive-sense RNA from a 100-nt template corresponding to the 3'-end of the viral antisense genome sequence, but that the RdRp could not synthesize the antisense genomic RNA from the template corresponding to the 5'-end of the positive-sense genome sequence. We also predicted that the 31 nt region at the 3'-end of the RNA antisense template forms a stem-loop structure. Deletion of this sequence resulted in the loss of complementary RNA synthesis by the RdRp, and connection of the 31 nt to the 3'-end of the inactive positive-sense RNA template resulted in the gain of complementary RNA synthesis by the RdRp. Similarly, an electrophoretic mobility shift assay further revealed that the RdRp bound to the antisense RNA specifically, but was dependent on the 31 nt at the 3'-end. Therefore, based on this observation and further deletion and mutation analyses, we concluded that the predicted stem-loop structure in the 31 nt end and the region close to the antisense viral genomic stem sequences are both important for initiating the positive-sense human norovirus genomic RNA synthesis by its RdRp.<br />Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.<br /> (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
- Subjects :
- Humans
Neoplasm Proteins genetics
Neoplasm Proteins metabolism
Norovirus genetics
RNA, Antisense genetics
RNA, Antisense metabolism
RNA, Viral biosynthesis
RNA, Viral genetics
RNA-Dependent RNA Polymerase genetics
RNA-Dependent RNA Polymerase metabolism
Genome, Viral
Neoplasm Proteins chemistry
Norovirus chemistry
Nucleic Acid Conformation
RNA, Antisense chemistry
RNA, Viral chemistry
RNA-Dependent RNA Polymerase chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1083-351X
- Volume :
- 297
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 34562449
- Full Text :
- https://doi.org/10.1016/j.jbc.2021.101225