Abstract
Core Qβ replicase comprises the Qβ virus–encoded RNA-dependent RNA polymerase (β-subunit) and the host Escherichia coli translational elongation factors EF-Tu and EF-Ts. The functions of the host proteins in the viral replicase are not clear. Structural analyses of RNA polymerization by core Qβ replicase reveal that at the initiation stage, the 3′-adenine of the template RNA provides a stable platform for de novo initiation. EF-Tu in Qβ replicase forms a template exit channel with the β-subunit. At the elongation stages, the C-terminal region of the β-subunit, assisted by EF-Tu, splits the temporarily double-stranded RNA between the template and nascent RNAs before translocation of the single-stranded template RNA into the exit channel. Therefore, EF-Tu in Qβ replicase modulates RNA elongation processes in a distinct manner from its established function in protein synthesis.
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Acknowledgements
We thank A. Hamada for technical assistance. We thank the beamline staff of BL-17A (KEK) for technical assistance during data collection. This work was supported by grants to K.T. from the Precursory Research for Embryonic Science and Technology program of the Japan Science and Technology Agency, the Funding program for Next Generation World-Leading Researchers (NEXT program) of the Japan Society for the Promotion of Science, the Takeda Science Foundation, the Mochida Memorial Foundation for Medical and Pharmaceutical Research and the Cell Science Research Foundation.
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K.T. planned and designed the research, K.T. and D.T. designed the experiments, D.T. conducted the experiments and D.T. and K.T. analyzed the data, discussed the results and wrote the paper.
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Supplementary Text and Figures
Supplementary Figures 1–4 and Supplementary Discussion (PDF 408 kb)
Supplementary Movie 1
Splitting the dsRNA of the template and growing RNAs. (MOV 1193 kb)
Supplementary Movie 2
The template entrance and exit channels in core Qβ replicase. (MOV 1122 kb)
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Takeshita, D., Tomita, K. Molecular basis for RNA polymerization by Qβ replicase. Nat Struct Mol Biol 19, 229–237 (2012). https://doi.org/10.1038/nsmb.2204
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DOI: https://doi.org/10.1038/nsmb.2204