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PMID: 19398578 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The NF90-NF45 complex functions as a negative regulator in the microRNA processing pathway.

Molecular and cellular biology ·Vol. 29 ·No. 13 ·2009-07-00 ·Pages 3754-69

Sakamoto S, Aoki K, Higuchi T, Todaka H, Morisawa K, Tamaki N, Hatano E, Fukushima A, Taniguchi T, Agata Y

Abstract

The positive regulatory machinery in the microRNA (miRNA) processing pathway is relatively well characterized, but negative regulation of the pathway is largely unknown. Here we show that a complex of nuclear factor 90 (NF90) and NF45 proteins functions as a negative regulator in miRNA biogenesis. Primary miRNA (pri-miRNA) processing into precursor miRNA (pre-miRNA) was inhibited by overexpression of the NF90 and NF45 proteins, and considerable amounts of pri-miRNAs accumulated in cells coexpressing NF90 and NF45. Treatment of cells overexpressing NF90 and NF45 with an RNA polymerase II inhibitor, alpha-amanitin, did not reduce the amounts of pri-miRNAs, suggesting that the accumulation of pri-miRNAs is not due to transcriptional activation. In addition, the NF90 and NF45 complex was not found to interact with the Microprocessor complex, which is a processing factor of pri-miRNAs, but was found to bind endogenous pri-miRNAs. NF90-NF45 exhibited higher binding activity for pri-let-7a than pri-miR-21. Of note, depletion of NF90 caused a reduction of pri-let-7a and an increase of mature let-7a miRNA, which has a potent antiproliferative activity, and caused growth suppression of transformed cells. These findings suggest that the association of the NF90-NF45 complex with pri-miRNAs impairs access of the Microprocessor complex to the pri-miRNAs, resulting in a reduction of mature miRNA production.

MeSH Terms
Cell Line Humans MicroRNAs/genetics,metabolism Multiprotein Complexes/metabolism Nuclear Factor 45 Protein/genetics,metabolism Nuclear Factor 90 Proteins/genetics,metabolism Proteins/genetics,metabolism RNA Interference RNA Precursors/genetics,metabolism RNA-Binding Proteins Recombinant Proteins/genetics,metabolism Ribonuclease III/genetics,metabolism
Chemicals
DGCR8 protein, human MicroRNAs Multiprotein Complexes Nuclear Factor 45 Protein Nuclear Factor 90 Proteins Proteins RNA Precursors RNA-Binding Proteins Recombinant Proteins DROSHA protein, human Ribonuclease III
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Sakamoto Shuji
Laboratory of Molecular Biology, Science Research Center, Kochi Medical School, Kochi 783-8505, Japan. sshuji@kochi-u.ac.jp
Aoki Kazuma
Higuchi Takuma
Todaka Hiroshi
Morisawa Keiko
Tamaki Nobuyuki
Hatano Etsuro
Fukushima Atsuki
Taniguchi Taketoshi
Agata Yasutoshi
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01836-08
Email
sshuji@kochi-u.ac.jp
Published
2009-07-00
Epub
2009-00-27
Pages
3754-69
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2698745
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.01836-08 PMC Full Text View journal details
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