DEAD-box RNA helicase subunits of the Drosha complex are required for processing of rRNA and a subset of microRNAs

Nature Cell Biology volume 9, pages604–611(2007)Cite this article

A Retraction to this article was published on 31 October 2014

Abstract

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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Figure 1: Embryonic and neonatal lethality of p68 and p72 knockout mice.
Figure 2: The role of RNA helicases p72 and p68 in miRNA or rRNA processing.
Figure 3: In vivo association of the Drosha complex with pri-miRNAs through p68–p72.
Figure 4: Biochemical isolation of mouse Drosha-containing complex.
Figure 5: Purified mDrosha complex mediates processing of miRNA and rRNA in vitro.

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Acknowledgements

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.).

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Author notes

  1. Toru Fukuda, Kaoru Yamagata and Sally Fujiyama: These authors contributed equally to this work.

Affiliations

  1. Institute of Molecular and Cellular Biosciences, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo, 113-0032, Japan

    Toru Fukuda, Kaoru Yamagata, Sally Fujiyama, Takahiro Matsumoto, Iori Koshida, Kimihiro Yoshimura, Masatomo Mihara, Takashi Nakamura, Chihiro Akimoto, Yoko Yamamoto, Shinichiro Takezawa, Hirochika Kitagawa, Ken-ichi Takeyama & Shigeaki Kato

  2. ERATO, Japan Science and Technology, Honcho 4-1-8, Kawaguchi, Saitama, 332-0012, Japan

    Kaoru Yamagata, Sally Fujiyama, Takahiro Matsumoto, Takashi Nakamura & Shigeaki Kato

  3. Applied Genomics, Molecular Medicine Labs., Drug Discovery Research, Astellas Pharma Inc., 21,Miyukigaoka, Tsukuba-shi, Ibaraki, 305-8585, Japan

    Masanori Naitou & Hideki Endoh

  4. Research Center for Genomic Medicine, Saitama Medical School, Yamane 1397-1, Hidakashi, Saitama, 350-1242, Japan

    Takenobu Katagiri

  5. Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, TX, USA

    Charles Foulds & Bert W. O'Malley

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  1. Toru Fukuda
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Contributions

The experimental concept was developed by S.K., B.W.O.M, S.T., K.T., H.E. and H.K. T.F., K.Y. and S.F. conducted most of experiments, and I.K., K.Y., M.M., M.N., T.N., C.A., Y.Y., T.K., C.F. and S.T. contributed materials and supported several experiments. S.K., T.F., K.Y. and S.F. wrote the manuscript.

Corresponding author

Correspondence to Shigeaki Kato.

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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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