A functional genomics screen for microRNA regulators of NF-kappaB signalingReportar como inadecuado




A functional genomics screen for microRNA regulators of NF-kappaB signaling - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

BMC Biology

, 11:19

First Online: 28 February 2013Received: 09 August 2012Accepted: 28 February 2013DOI: 10.1186-1741-7007-11-19

Cite this article as: Olarerin-George, A.O., Anton, L., Hwang, YC. et al. BMC Biol 2013 11: 19. doi:10.1186-1741-7007-11-19

Abstract

BackgroundThe nuclear factor-KappaB NF-κB pathway is conserved from fruit flies to humans and is a key mediator of inflammatory signaling. Aberrant regulation of NF-κB is associated with several disorders including autoimmune disease, chronic inflammation, and cancer, making the NF-κB pathway an attractive therapeutic target. Many regulatory components of the NF-κB pathway have been identified, including microRNAs miRNAs. miRNAs are small non-coding RNAs and are common components of signal transduction pathways. Here we present a cell-based functional genomics screen to systematically identify miRNAs that regulate NF-κB signaling.

ResultsWe screened a library of miRNA mimics using a NF-κB reporter cell line in the presence and absence of tumor necrosis factor +- TNF. There were 9 and 15 hits in the -TNF and +TNF screens, respectively. We identified putative functional targets of these hits by integrating computational predictions with NF-κB modulators identified in a previous genome-wide cDNA screen. miR-517a and miR-517c were the top hits, activating the reporter 86- and 126-fold, respectively. Consistent with these results, miR-517a-c induced the expression of endogenous NF-κB targets and promoted the nuclear localization of p65 and the degradation of IκB. We identified TNFAIP3 interacting protein1 TNIP1 as a target and characterized a functional SNP in the miR-517a-c binding site. Lastly, miR-517a-c induced apoptosis in vitro, which was phenocopied by knockdown of TNIP1.

ConclusionsOur study suggests that miRNAs are common components of NF-κB signaling and miR-517a-c may play an important role in linking NF-κB signaling with cell survival through TNIP1.

KeywordsmicroRNA miRNA miR-517 NF-kappaB TNIP1 TNF RNAi screen apoptosis Abbreviationsbpbase pair

CASP3caspase 3

CASP7caspase 7

CScontext score

DMEMDulbecco-s modified Eagle-s medium

EDTAethylenediaminetetraacetic acid

ELISAenzyme-linked immunosorbent assay

FBSfetal bovine serum

HRPhorseradish peroxidase

HUVECshuman umbilical vein endothelial cells

IκBI kappa-B

IgGimmunoglobulin G

IKKI kappa-B kinase

ILinterleukin

LPSlipopolysaccharide

madmedian absolute deviations

miRNAsmicro RNAs

ncRNAsnon-coding RNAs

NF-κBnuclear factor kappa-B

NF-κB-lucnuclear factor kappa-B responsive luciferase

NKRFnuclear factor kappa-B repressing factor

PARP1polyADP-ribose polymerase 1

PCRpolymerase chain reaction

siRNAssmall interfering RNAs

SNPsingle nucleotide polymorphism

TETris-ethylenediaminetetraacetic acid

TNFtumor necrosis factor

TNIP1tumor necrosis factor alpha-induced protein 3 interacting protein

UTRuntranslated region.

Electronic supplementary materialThe online version of this article doi:10.1186-1741-7007-11-19 contains supplementary material, which is available to authorized users.

Download fulltext PDF



Autor: Anthony O Olarerin-George - Lauren Anton - Yih-Chii Hwang - Michal A Elovitz - John B Hogenesch

Fuente: https://link.springer.com/







Documentos relacionados