MicroRNAs (miRNAs) control cell proliferation, differentiation and fate through modulation of gene expression by partially base-pairing with target mRNA sequences1,2,3,4,5,6. Drosha is an RNase III enzyme that is the catalytic subunit of a large complex that cleaves pri-miRNAs with distinct structures into pre-miRNAs7. Here, we show that both the p68 and p72 DEAD-box RNA helicase subunits8,9,10 in the mouse Drosha complex are indispensable for survival in mice, and both are required for primary miRNA and rRNA processing. Gene disruption of either p68 or p72 in mice resulted in early lethality, and in both p68−/− and p72−/− embryos, expression levels of a set of, but not all, miRNAs and 5.8S rRNA were significantly lowered. In p72−/− MEF cells, expression of p72, but not a mutant lacking ATPase activity, restored the impaired expression of miRNAs and 5.8S rRNA. Furthermore, we purified the large complex of mouse Drosha and showed it could generate pre-miRNA and 5.8S rRNA in vitro. Thus, we suggest that DEAD-box RNA helicase subunits are required for recognition of a subset of primary miRNAs in mDrosha-mediated processing.
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We thank T. Watanabe, H. Shiina, J. Miyamoto, K. Sekine, R. Fujiki, Y. Imai and S. Tanaka for generation of knockout mice, and H. Higuchi for manuscript preparation. This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) and priority areas from the Ministry of Education, Culture, Sports, Science and Technology (to S.K.).
The authors declare no competing financial interests.
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Fukuda, T., Yamagata, K., Fujiyama, S. et al. DEAD-box RNA helicase subunits of the Drosha complex are required for processing of rRNA and a subset of microRNAs. Nat Cell Biol 9, 604–611 (2007). https://doi.org/10.1038/ncb1577
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