Long noncoding RNA Saf and splicing factor 45 increase soluble Fas and resistance to apoptosis

Oncotarget
Olga VillamizarAndrew Wilber

Abstract

In multicellular organisms, cell growth and differentiation is controlled in part by programmed cell death or apoptosis. One major apoptotic pathway is triggered by Fas receptor (Fas)-Fas ligand (FasL) interaction. Neoplastic cells are frequently resistant to Fas-mediated apoptosis, evade Fas signals through down regulation of Fas and produce soluble Fas proteins that bind FasL thereby blocking apoptosis. Soluble Fas (sFas) is an alternative splice product of Fas pre-mRNA, commonly created by exclusion of transmembrane spanning sequences encoded within exon 6 (FasΔEx6). Long non-coding RNAs (lncRNAs) interact with other RNAs, DNA, and proteins to regulate gene expression. One lncRNA, Fas-antisense or Saf, was shown to participate in alternative splicing of Fas pre-mRNA through unknown mechanisms. We show that Saf is localized in the nucleus where it interacts with Fas receptor pre-mRNA and human splicing factor 45 (SPF45) to facilitate alternative splicing and exclusion of exon 6. The product is a soluble Fas protein that protects cells against FasL-induced apoptosis. Collectively, these studies reveal a novel mechanism to modulate this critical cell death program by an lncRNA and its protein partner.

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Methods Mentioned

BETA
fluorescence-activated cell sorting
PCR
flow cytometry
ELISA
FACS
pull-down
pulldown
co-immunoprecipitation
immunoprecipitation
in

Software Mentioned

GraphPad
IntaRNA
Prism
Excel
FlowJo
ImageJ
Protein Prospector
SEQUEST
Mascot
CellQuest

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