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Beyond Secondary Structure: Primary-Sequence Determinants License Pri-miRNA Hairpins for Processing

Author(s)
Auyeung, Vincent C.; Ulitsky, Igor; McGeary, Sean Edward; Bartel, David; Auyeung, Vincent C.
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Abstract
To use microRNAs to downregulate mRNA targets, cells must first process these ∼22 nt RNAs from primary transcripts (pri-miRNAs). These transcripts form RNA hairpins important for processing, but additional determinants must distinguish pri-miRNAs from the many other hairpin-containing transcripts expressed in each cell. Illustrating the complexity of this recognition, we show that most Caenorhabditis elegans pri-miRNAs lack determinants required for processing in human cells. To find these determinants, we generated many variants of four human pri-miRNAs, sequenced millions that retained function, and compared them with the starting variants. Our results confirmed the importance of pairing in the stem and revealed three primary-sequence determinants, including an SRp20-binding motif (CNNC) found downstream of most pri-miRNA hairpins in bilaterian animals, but not in nematodes. Adding this and other determinants to C. elegans pri-miRNAs imparted efficient processing in human cells, thereby confirming the importance of primary-sequence determinants for distinguishing pri-miRNAs from other hairpin-containing transcripts.
Date issued
2013-02
URI
http://hdl.handle.net/1721.1/101253
Department
Harvard University--MIT Division of Health Sciences and Technology; Massachusetts Institute of Technology. Department of Biology; Whitehead Institute for Biomedical Research
Journal
Cell
Publisher
Elsevier
Citation
Auyeung, Vincent C., Igor Ulitsky, Sean E. McGeary, and David P. Bartel. “Beyond Secondary Structure: Primary-Sequence Determinants License Pri-miRNA Hairpins for Processing.” Cell 152, no. 4 (February 2013): 844–858.
Version: Author's final manuscript
ISSN
00928674
1097-4172

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