DEAD-box RNA helicase subunits of the Drosha complex are required for processing of rRNA and a subset of microRNAs.
MicroRNAs (miRNAs) control cell proliferation, differentiation and fate through modulation of gene expression by partially base-pairing with target mRNA sequences. Drosha is an RNase III enzyme that is the catalytic subunit of a large complex that cleaves pri-miRNAs with distinct structures into pre-miRNAs. Here, we show that both the p68 and p72 DEAD-box RNA helicase subunits in the mouse Drosha complex are indispensable for survival in mice, and both are required for primary miRNA and rRNA processing. Gene disruption of either p68 or p72 in mice resulted in early lethality, and in both p68(-/-) and p72(-/-) embryos, expression levels of a set of, but not all, miRNAs and 5.8S rRNA were significantly lowered. In p72(-/-) MEF cells, expression of p72, but not a mutant lacking ATPase activity, restored the impaired expression of miRNAs and 5.8S rRNA. Furthermore, we purified the large complex of mouse Drosha and showed it could generate pre-miRNA and 5.8S rRNA in vitro. Thus, we suggest that DEAD-box RNA helicase subunits are required for recognition of a subset of primary miRNAs in mDrosha-mediated processing.
Fukuda
Toru
T
Institute of Molecular and Cellular Biosciences, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-0032, Japan.
Yamagata
Kaoru
K
Fujiyama
Sally
S
Matsumoto
Takahiro
T
Koshida
Iori
I
Yoshimura
Kimihiro
K
Mihara
Masatomo
M
Naitou
Masanori
M
Endoh
Hideki
H
Nakamura
Takashi
T
Akimoto
Chihiro
C
Yamamoto
Yoko
Y
Katagiri
Takenobu
T
Foulds
Charles
C
Takezawa
Shinichiro
S
Kitagawa
Hirochika
H
Takeyama
Ken-ichi
K
O'Malley
Bert W
BW
Kato
Shigeaki
S
D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D053487:DEAD-box RNA Helicases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D012322:RNA Precursors / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000378:metabolism*; D012340:RNA, Ribosomal, 5.8S / Q000378:metabolism; D043244:Ribonuclease III / Q000378:metabolism*
D007527:Isoenzymes; D035683:MicroRNAs; D012322:RNA Precursors; D012335:RNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S; C513400:Drosha protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases