Abstract
The evolutionary origin of human hepatitis delta virus (HDV) replication by RNA-directed transcription is unclear. Here we identify two species of 5′-capped, ∼18–25-nucleotide small RNAs. One was of antigenomic polarity, corresponding to the 5′ end of hepatitis delta antigen (HDAg) mRNA, and interacted with HDAg and RNA polymerase II (Pol II), whereas the other mapped to a structurally analogous region on the genomic RNA hairpin. An HDAg-interaction screen indicated that HDAg interacts with MOV10, the human homolog of the Arabidopsis thaliana RNA amplification factor gene SDE3 and Drosophila melanogaster RISC-maturation factor gene Armitage (armi). MOV10 knockdown inhibited HDV replication, but not HDAg mRNA translation, further supporting a role for MOV10 in RNA-directed transcription. Together, our studies define RNA hairpins as critical elements for the initiation of HDV-related, RNA-directed transcription. The identification of capped small RNAs and the involvement of MOV10 in HDV replication further suggest a conserved mechanism related to RNA-directed transcription in lower eukaryotes.
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Change history
22 June 2008
In the version of this article initially published online, the top panel in Figure 3d was mistakenly replaced with a panel from Figure 3c. The error has been corrected for all versions of this article.
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Acknowledgements
We thank J. Glenn (Stanford University) for reagents, R. Lanford (Southwest Foundation for Biomedical Research, San Antonio) for HDV virions and A. Guzzetta (Stanford University) for mass spectrometry. This work was supported by grants DK78424 and AI71068 from the National Institutes of Health (to M.A.K.), a Helen Hay Whitney Foundation Postdoctoral Research Fellowship to D.C. and a Stanford Dean's Postdoctoral Fellowship to D.H.
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D.H. designed and performed the majority of the experiments. D.C. did the proteomic screens and contributed to the immunoprecipitation experiments. Y.H. assisted D.H. in performing the experiments. P.P. did the small RNA sequencing. A.Z.F. provided input on sequencing strategies and suggestions in experimental design. M.A.K. served as supervisor and provided scientific input to experimental design and data interpretation. D.H. wrote the manuscript with input from D.C., P.P., A.Z.F. and M.A.K. All authors approved the final manuscript.
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Supplementary Figures 1–3, Supplementary Table 1 and Supplementary Methods (PDF 409 kb)
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Haussecker, D., Cao, D., Huang, Y. et al. Capped small RNAs and MOV10 in human hepatitis delta virus replication. Nat Struct Mol Biol 15, 714–721 (2008). https://doi.org/10.1038/nsmb.1440
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DOI: https://doi.org/10.1038/nsmb.1440
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