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.
authors.affiliation: Center for RNA Research, Institute for Basic Science, Seoul 151-742, Korea; School of Biological Sciences, Seoul National University, Seoul 151-742, Korea.
authors.forename: Tuan Anh
authors.initials: TA
authors.lastname: Nguyen
authors.affiliation: Department of Physics and Astronomy, Seoul National University, Seoul 151-742, Korea; National Center for Creative Research Initiatives, Seoul National University, Seoul 151-742, Korea; Institute of Applied Physics, Seoul National University, Seoul 151-742, Korea.
authors.forename: Myung Hyun
authors.initials: MH
authors.lastname: Jo
authors.affiliation: Center for RNA Research, Institute for Basic Science, Seoul 151-742, Korea; School of Biological Sciences, Seoul National University, Seoul 151-742, Korea.
authors.forename: Yeon-Gil
authors.initials: YG
authors.lastname: Choi
authors.affiliation: Center for RNA Research, Institute for Basic Science, Seoul 151-742, Korea; School of Biological Sciences, Seoul National University, Seoul 151-742, Korea.
authors.forename: Joha
authors.initials: J
authors.lastname: Park
authors.affiliation: Center for RNA Research, Institute for Basic Science, Seoul 151-742, Korea; School of Biological Sciences, Seoul National University, Seoul 151-742, Korea.
authors.forename: S Chul
authors.initials: SC
authors.lastname: Kwon
authors.affiliation: Department of Physics and Astronomy, Seoul National University, Seoul 151-742, Korea; National Center for Creative Research Initiatives, Seoul National University, Seoul 151-742, Korea; Institute of Applied Physics, Seoul National University, Seoul 151-742, Korea; Department of Biophysics and Chemical Biology, Seoul National University, Seoul 151-742, Korea.
authors.forename: Sungchul
authors.initials: S
authors.lastname: Hohng
authors.affiliation: Center for RNA Research, Institute for Basic Science, Seoul 151-742, Korea; School of Biological Sciences, Seoul National University, Seoul 151-742, Korea. Electronic address: narrykim@snu.ac.kr.
authors.forename: V Narry
authors.initials: VN
authors.lastname: Kim
authors.affiliation: Center for RNA Research, Institute for Basic Science, Seoul 151-742, Korea; School of Biological Sciences, Seoul National University, Seoul 151-742, Korea. Electronic address: jaesungwoo@snu.ac.kr.
authors.forename: Jae-Sung
authors.initials: JS
authors.lastname: Woo
mesh_terms: D001483:Base Sequence; D019281:Dimerization; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D059372:Nucleotide Motifs; D012323:RNA Processing, Post-Transcriptional; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D043244:Ribonuclease III / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C474043:DGCR8 protein, human; D035683:MicroRNAs; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C517918:DROSHA protein, human; D043244:Ribonuclease III