Functional anatomy of the human microprocessor

Tuan Anh Nguyen; Myung Hyun Jo; Yeon Gil Choi; Joha Park; S. Chul Kwon; Sungchul Hohng; V. Narry Kim; Jae Sung Woo

doi:10.1016/j.cell.2015.05.010

Functional anatomy of the human microprocessor

Tuan Anh Nguyen, Myung Hyun Jo, Yeon Gil Choi, Joha Park, S. Chul Kwon, Sungchul Hohng, V. Narry Kim, Jae Sung Woo

Research output: Contribution to journal › Article › peer-review

284 Citations (Scopus)

Abstract

MicroRNA (miRNA) maturation is initiated by Microprocessor composed of RNase III DROSHA and its cofactor DGCR8, whose fidelity is critical for generation of functional miRNAs. To understand how Microprocessor recognizes pri-miRNAs, we here reconstitute human Microprocessor with purified recombinant proteins. We find that Microprocessor is an ∼ 364 kDa heterotrimeric complex of one DROSHA and two DGCR8 molecules. Together with a 23-amino acid peptide from DGCR8, DROSHA constitutes a minimal functional core. DROSHA serves as a "ruler" by measuring 11 bp from the basal ssRNA-dsRNA junction. DGCR8 interacts with the stem and apical elements through its dsRNA-binding domains and RNA-binding heme domain, respectively, allowing efficient and accurate processing. DROSHA and DGCR8, respectively, recognize the basal UG and apical UGU motifs, which ensure proper orientation of the complex. These findings clarify controversies over the action mechanism of DROSHA and allow us to build a general model for pri-miRNA processing.

Original language	English
Pages (from-to)	1374-1387
Number of pages	14
Journal	Cell
Volume	161
Issue number	6
DOIs	https://doi.org/10.1016/j.cell.2015.05.010
Publication status	Published - 2015 Jun 5
Externally published	Yes

Bibliographical note

Funding Information:
We are grateful to members of our laboratory for discussion and technical help, especially Eunhye Shin, Eunji Kim, Sunah Kim, and Ahyoung Cho for their technical assistance. We thank Dr. Yeon-Soo Seo for the gift of the pET-28a plasmid. This work was supported by IBS-R008-D1 of Institute for Basic Science from the Ministry of Science, ICT, and Future Planning of Korea (T.A.N., Y.-G.C., J.P., S.C.K., J.-S.W., and V.N.K.), the BK21 Research Fellowships from the Ministry of Education of Korea (J.P.), and Creative Research Initiatives (2009-0081562) of the National Research Foundation of Korea (M.H.J. and S.H.).

Publisher Copyright:
© 2015 Elsevier Inc.

ASJC Scopus subject areas

General Biochemistry,Genetics and Molecular Biology

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10.1016/j.cell.2015.05.010

Cite this

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title = "Functional anatomy of the human microprocessor",

abstract = "MicroRNA (miRNA) maturation is initiated by Microprocessor composed of RNase III DROSHA and its cofactor DGCR8, whose fidelity is critical for generation of functional miRNAs. To understand how Microprocessor recognizes pri-miRNAs, we here reconstitute human Microprocessor with purified recombinant proteins. We find that Microprocessor is an ∼ 364 kDa heterotrimeric complex of one DROSHA and two DGCR8 molecules. Together with a 23-amino acid peptide from DGCR8, DROSHA constitutes a minimal functional core. DROSHA serves as a {"}ruler{"} by measuring 11 bp from the basal ssRNA-dsRNA junction. DGCR8 interacts with the stem and apical elements through its dsRNA-binding domains and RNA-binding heme domain, respectively, allowing efficient and accurate processing. DROSHA and DGCR8, respectively, recognize the basal UG and apical UGU motifs, which ensure proper orientation of the complex. These findings clarify controversies over the action mechanism of DROSHA and allow us to build a general model for pri-miRNA processing.",

author = "Nguyen, {Tuan Anh} and Jo, {Myung Hyun} and Choi, {Yeon Gil} and Joha Park and Kwon, {S. Chul} and Sungchul Hohng and Kim, {V. Narry} and Woo, {Jae Sung}",

note = "Funding Information: We are grateful to members of our laboratory for discussion and technical help, especially Eunhye Shin, Eunji Kim, Sunah Kim, and Ahyoung Cho for their technical assistance. We thank Dr. Yeon-Soo Seo for the gift of the pET-28a plasmid. This work was supported by IBS-R008-D1 of Institute for Basic Science from the Ministry of Science, ICT, and Future Planning of Korea (T.A.N., Y.-G.C., J.P., S.C.K., J.-S.W., and V.N.K.), the BK21 Research Fellowships from the Ministry of Education of Korea (J.P.), and Creative Research Initiatives (2009-0081562) of the National Research Foundation of Korea (M.H.J. and S.H.). Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

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AU - Kwon, S. Chul

AU - Hohng, Sungchul

AU - Kim, V. Narry

AU - Woo, Jae Sung

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PY - 2015/6/5

Y1 - 2015/6/5

N2 - MicroRNA (miRNA) maturation is initiated by Microprocessor composed of RNase III DROSHA and its cofactor DGCR8, whose fidelity is critical for generation of functional miRNAs. To understand how Microprocessor recognizes pri-miRNAs, we here reconstitute human Microprocessor with purified recombinant proteins. We find that Microprocessor is an ∼ 364 kDa heterotrimeric complex of one DROSHA and two DGCR8 molecules. Together with a 23-amino acid peptide from DGCR8, DROSHA constitutes a minimal functional core. DROSHA serves as a "ruler" by measuring 11 bp from the basal ssRNA-dsRNA junction. DGCR8 interacts with the stem and apical elements through its dsRNA-binding domains and RNA-binding heme domain, respectively, allowing efficient and accurate processing. DROSHA and DGCR8, respectively, recognize the basal UG and apical UGU motifs, which ensure proper orientation of the complex. These findings clarify controversies over the action mechanism of DROSHA and allow us to build a general model for pri-miRNA processing.

AB - MicroRNA (miRNA) maturation is initiated by Microprocessor composed of RNase III DROSHA and its cofactor DGCR8, whose fidelity is critical for generation of functional miRNAs. To understand how Microprocessor recognizes pri-miRNAs, we here reconstitute human Microprocessor with purified recombinant proteins. We find that Microprocessor is an ∼ 364 kDa heterotrimeric complex of one DROSHA and two DGCR8 molecules. Together with a 23-amino acid peptide from DGCR8, DROSHA constitutes a minimal functional core. DROSHA serves as a "ruler" by measuring 11 bp from the basal ssRNA-dsRNA junction. DGCR8 interacts with the stem and apical elements through its dsRNA-binding domains and RNA-binding heme domain, respectively, allowing efficient and accurate processing. DROSHA and DGCR8, respectively, recognize the basal UG and apical UGU motifs, which ensure proper orientation of the complex. These findings clarify controversies over the action mechanism of DROSHA and allow us to build a general model for pri-miRNA processing.

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Functional anatomy of the human microprocessor

Abstract

Bibliographical note

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