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The Microprocessor complex mediates the genesis of microRNAs

Nature volume 432, pages 235–240 (2004)Cite this article

Abstract

MicroRNAs (miRNAs) are a growing family of small non-protein-coding regulatory genes that regulate the expression of homologous target-gene transcripts. They have been implicated in the control of cell death and proliferation in flies1,2, haematopoietic lineage differentiation in mammals3, neuronal patterning in nematodes4 and leaf and flower development in plants5,6,7,8. miRNAs are processed by the RNA-mediated interference machinery. Drosha is an RNase III enzyme that was recently implicated in miRNA processing. Here we show that human Drosha is a component of two multi-protein complexes. The larger complex contains multiple classes of RNA-associated proteins including RNA helicases, proteins that bind double-stranded RNA, novel heterogeneous nuclear ribonucleoproteins and the Ewing's sarcoma family of proteins. The smaller complex is composed of Drosha and the double-stranded-RNA-binding protein, DGCR8, the product of a gene deleted in DiGeorge syndrome. In vivo knock-down and in vitro reconstitution studies revealed that both components of this smaller complex, termed Microprocessor, are necessary and sufficient in mediating the genesis of miRNAs from the primary miRNA transcript.

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Figure 1: Isolation of Drosha-containing complexes.
Figure 2: Composition and miRNA processing activity of Superose 6 gel-filtration fractions.
Figure 3: Microprocessor is the Drosha–DGCR8 complex mediating miRNA processing.
Figure 4: Reconstitution of Microprocessor by using recombinant Drosha and DGCR8.
Figure 5: A role for Microprocessor in vivo in miRNA processing.

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References

  1. Brennecke, J. et al. bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113, 25–36 (2003)

    Article  CAS  Google Scholar 

  2. Xu, P. et al. The Drosophila microRNA mir-14 suppresses cell death and is required for normal fat metabolism. Curr. Biol. 13, 790–795 (2003)

    Article  CAS  Google Scholar 

  3. Chen, C. Z. et al. MicroRNAs modulate hematopoietic lineage differentiation. Science 303, 83–86 (2004)

    Article  ADS  CAS  Google Scholar 

  4. Johnston, R. J. & Hobert, O. A microRNA controlling left/right neuronal asymmetry in Caenorhabditis elegans. Nature 426, 845–849 (2003)

    Article  ADS  CAS  Google Scholar 

  5. Aukerman, M. J. & Sakai, H. Regulation of flowering time and floral organ identity by a microRNA and its APETALA2-like target genes. Plant Cell 15, 2730–2741 (2003)

    Article  CAS  Google Scholar 

  6. Chen, X. et al. A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development. Science 303, 2022–2025 (2004)

    Article  ADS  CAS  Google Scholar 

  7. Emery, J. F. et al. Radial patterning of Arabidopsis shoots by class III HD-ZIP and KANADI genes. Curr. Biol. 13, 1768–1774 (2003)

    Article  CAS  Google Scholar 

  8. Palatnik, J. F. et al. Control of leaf morphogenesis by microRNAs. Nature 425, 257–263 (2003)

    Article  ADS  CAS  Google Scholar 

  9. Lagos-Quintana, M. et al. Identification of novel genes coding for small expressed RNAs. Science 294, 853–858 (2001)

    Article  ADS  CAS  Google Scholar 

  10. Lau, N. C. et al. An abundant class of tiny RNAs with probable regulatory roles in Caenorhabditis elegans. Science 294, 858–862 (2001)

    Article  ADS  CAS  Google Scholar 

  11. Lee, R. C. & Ambros, V. An extensive class of small RNAs in Caenorhabditis elegans. Science 294, 862–864 (2001)

    Article  ADS  CAS  Google Scholar 

  12. Aravin, A. A. et al. The small RNA profile during Drosophila melanogaster development. Dev. Cell 5, 337–350 (2003)

    Article  CAS  Google Scholar 

  13. Lagos-Quintana, M. et al. New microRNAs from mouse and human. RNA 9, 175–179 (2003)

    Article  CAS  Google Scholar 

  14. Lim, L. P. et al. The microRNAs of Caenorhabditis elegans. Genes Dev. 17, 991–1008 (2003)

    Article  CAS  Google Scholar 

  15. Lee, Y., Jeon, K., Lee, J. T., Kim, S. & Kim, V. N. MicroRNA maturation: stepwise processing and subcellular localization. EMBO J. 21, 4663–4670 (2002)

    Article  CAS  Google Scholar 

  16. Zeng, Y. & Cullen, B. R. MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms. Proc. Natl Acad. Sci. USA 100, 9779–9784 (2003)

    Article  ADS  CAS  Google Scholar 

  17. Lee, Y. et al. The nuclear RNase III Drosha initiates microRNA processing. Nature 425, 415–419 (2003)

    Article  ADS  CAS  Google Scholar 

  18. Denali, A. M., Tops, B. B. J., Plasterk, R. H. A., Ketting, R. F. & Hannon, G. J. Processing of primary microRNAs by the Microprocessor complex. Nature doi:10.1038/nature03049 (this issue)

  19. Yamagishi, H. & Srivastava, D. Unraveling the genetic and developmental mysteries of 22q11 deletion syndrome. Trends Mol. Med. 9, 383–389 (2003)

    Article  CAS  Google Scholar 

  20. Shiohama, A., Sasaki, T., Noda, S., Minoshima, S. & Shimizu, N. Molecular cloning and expression analysis of a novel gene DGCR8 located in DiGeorge syndrome chromosomal region. Biochem. Biophys. Res. Commun. 304, 184–190 (2003)

    Article  CAS  Google Scholar 

  21. Arvand, A. & Denny, C. T. Biology of EWS/ETS fusions in Ewing's family tumors. Oncogene 20, 5747–5754 (2001)

    Article  CAS  Google Scholar 

  22. Wu, H., Xu, H., Miraglia, L. J. & Crooke, S. T. Human RNase III is a 160-kDa protein involved in preribosomal RNA processing. J. Biol. Chem. 275, 36957–36965 (2000)

    Article  CAS  Google Scholar 

  23. Bochar, D. A. et al. BRCA1 is associated with a human SWI/SNF-related complex: linking chromatin remodeling to breast cancer. Cell 102, 257–265 (2000)

    Article  CAS  Google Scholar 

  24. Dong, Y. et al. Regulation of BRCC, a holoenzyme complex containing BRCA1 and BRCA2, by a signalosome-like subunit and its role in DNA repair. Mol. Cell 12, 1087–1099 (2003)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank V. N. Kim for the Drosha cDNA; O. Delattre and A. I. Lamond for the gift of EWS and DDX17 antibodies, respectively; K. Nishikura for providing recombinant Drosha; and T. Beer (Wistar Proteomocs Facility) for expertise in the microcapillary HPLC/mass spectrometry. R.S. was supported by grants from the NIH and the American Cancer Institute. R.G. is a fellow of the Jane Coffin Child Memorial Fund for Medical Research.

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  1. Richard I. Gregory and Kai-ping Yan: These authors contributed equally to this work

Authors and Affiliations

  1. The Wistar Institute, 3601 Spruce Street, Philadelphia, Pennsylvania, 19104, USA

    Richard I. Gregory, Kai-ping Yan, Govindasamy Amuthan, Thimmaiah Chendrimada, Behzad Doratotaj, Neil Cooch & Ramin Shiekhattar

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  1. Richard I. Gregory
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Correspondence to Ramin Shiekhattar.

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Supplementary information

Supplementary Figure 1

Diagrammatic depiction of Drosha-associated proteins and their domain structures. (PDF 47 kb)

Supplementary Figure 2

The bulk of miRNA processing activity resides in Drosha/DGCR8 complex. (PDF 281 kb)

Supplementary Figure 3

Analysis of miRNA precursors. (PDF 425 kb)

Supplementary Figure Legends (DOC 20 kb)

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Gregory, R., Yan, Kp., Amuthan, G. et al. The Microprocessor complex mediates the genesis of microRNAs. Nature 432, 235–240 (2004). https://doi.org/10.1038/nature03120

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