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Non-Coding RNA 2016, 2(4), 12; doi:10.3390/ncrna2040012

TWEAK Negatively Regulates Human Dicer

1
CHUQ Research Center/CHUL, 2705 Blvd Laurier, Quebec, QC G1V 4G2, Canada
2
Department of Microbiology-Infectious Disease and Immunity and Faculty of Medicine, Université Laval, Quebec, QC G1V 0A6, Canada
The first two authors should be regarded as joint First Authors.
*
Author to whom correspondence should be addressed.
Academic Editor: George A. Calin
Received: 7 September 2016 / Revised: 7 October 2016 / Accepted: 14 October 2016 / Published: 25 October 2016
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Abstract

The ribonuclease Dicer plays a central role in the microRNA pathway by processing microRNA precursors (pre-microRNAs) into microRNAs, a class of 19- to 24-nucleotide non-coding RNAs that regulate expression of ≈60% of the genes in humans. To gain further insights into the function and regulation of Dicer in human cells, we performed a yeast two-hybrid (Y2HB) screen using human Dicer double-stranded RNA-binding domain (dsRBD) as bait. This approach identified tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) as a Dicer-interacting protein candidate. Confocal immunofluorescence microscopy revealed the colocalization of Dicer and TWEAK proteins at the perinuclear region of HeLa cells. The Dicer-TWEAK protein interaction was confirmed by coimmunoprecipitation and found not likely to be mediated by RNA. TWEAK dose-dependently reduced pre-microRNA conversion into mature microRNA in Dicer activity assays using extracts of transfected human HEK 293 cells. TWEAK expression also impaired microRNA-guided RNA silencing of a reporter gene induced by a pre-microRNA. These findings suggest a role for TWEAK—a pro-inflammatory cytokine—in regulating Dicer function and microRNA biogenesis, and its possible involvement in regulating gene expression during inflammatory processes and diseases. View Full-Text
Keywords: yeast two-hybrid system; Dicer; TWEAK; protein interaction; microRNA yeast two-hybrid system; Dicer; TWEAK; protein interaction; microRNA
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Lambert, M.; Pépin, G.; Peralta-Zaragoza, O.; Matusiak, R.; Ly, S.; Landry, P.; Provost, P. TWEAK Negatively Regulates Human Dicer. Non-Coding RNA 2016, 2, 12.

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