한빛사 논문
S. Chul Kwon,1,2 S. Chan Baek,1,2 Yeon-Gil Choi,1,2 Jihye Yang,1,2 Young-suk Lee,1,2 Jae-Sung Woo,1,3,* and V. Narry Kim1,2,4,*
1 Center for RNA Research, Institute for Basic Science, Seoul 08826, Korea
2 School of Biological Sciences, Seoul National University, Seoul 08826, Korea
3 Department of Life Sciences, Korea University, Seoul 02841, Korea
4 Lead Contact
*Correspondence : Jae-Sung Woo, V. Narry Kim
Abstract
Microprocessor, composed of DROSHA and its cofactor DGCR8, initiates microRNA (miRNA) biogenesis by processing the primary transcripts of miRNA (pri-miRNAs). Here we investigate the mechanism by which Microprocessor selects the cleavage site with single-nucleotide precision, which is crucial for the specificity and functionality of miRNAs. By testing ∼40,000 pri-miRNA variants, we find that for some pri-miRNAs the cleavage site is dictated mainly by the mGHG motif embedded in the lower stem region of pri-miRNA. Structural modeling and deep-sequencing-based complementation experiments show that the double-stranded RNA-binding domain (dsRBD) of DROSHA recognizes mGHG to place the catalytic center in the appropriate position. The mGHG motif as well as the mGHG-recognizing residues in DROSHA dsRBD are conserved across eumetazoans, suggesting that this mechanism emerged in an early ancestor of the animal lineage. Our findings provide a basis for the understanding of miRNA biogenesis and rational design of accurate small-RNA-based gene silencing.
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