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Re-evaluation of the roles of DROSHA, Exportin 5, and DICER in microRNA biogenesis

Cited 168 time in webofscience Cited 163 time in scopus
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Title
Re-evaluation of the roles of DROSHA, Exportin 5, and DICER in microRNA biogenesis
Author(s)
Young-Kook Kim; Boseon Kim; V. Narry Kim
Subject
microRNA, ; DROSHA, ; Exportin 5, ; DICER, ; knockout
Publication Date
2016-03
Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.113, no.13, pp.E1881 - E1889
Publisher
NATL ACAD SCIENCES
Abstract
Biogenesis of canonical microRNAs (miRNAs) involves multiple steps: nuclear processing of primary miRNA (pri-miRNA) by DROSHA, nuclear export of precursor miRNA (pre-miRNA) by Exportin 5 (XPO5), and cytoplasmic processing of pre-miRNA by DICER. To gain a deeper understanding of the contribution of each of these maturation steps, we deleted DROSHA, XPO5, and DICER in the same human cell line, and analyzed their effects on miRNA biogenesis. Canonical miRNA production was completely abolished in DROSHA-deleted cells, whereas we detected a few DROSHA-independent miRNAs including three previously unidentified noncanonical miRNAs (miR-7706, miR-3615, and miR-1254). In contrast to DROSHA knockout, many canonical miRNAs were still detected without DICER albeit at markedly reduced levels. In the absence of DICER, pre-miRNAs are loaded directly onto AGO and trimmed at the 3' end, yielding miRNAs from the 5' strand (5p miRNAs). Interestingly, in XPO5 knockout cells, most miRNAs are affected only modestly, suggesting that XPO5 is necessary but not critical for miRNA maturation. Our study demonstrates an essential role of DROSHA and an important contribution of DICER in the canonical miRNA pathway, and reveals that the function of XPO5 can be complemented by alternative mechanisms. Thus, this study allows us to understand differential contributions of key biogenesis factors, and provides with valuable resources for miRNA research.
URI
https://pr.ibs.re.kr/handle/8788114/2512
DOI
10.1073/pnas.1602532113
ISSN
0027-8424
Appears in Collections:
Center for RNA Research(RNA 연구단) > 1. Journal Papers (저널논문)
Files in This Item:
PNAS-2016-Kim-E1881-9.pdfDownload

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