TY - JOUR
T1 - Intracellular calcium regulates nonsense-mediated mRNA decay
AU - Nickless, Andrew
AU - Jackson, Erin
AU - Marasa, Jayne
AU - Nugent, Patrick
AU - Mercer, Robert W.
AU - Piwnica-Worms, David
AU - You, Zhongsheng
N1 - Funding Information:
We thank P. Wu and X. Chen for their contributions in the early stage of this project. We are grateful to O. Mühlemann (University of Bern) for providing the pβ510 reporter construct, which we used to obtain the TRB minigene reporter sequence for our reporter construction, and R. Mecham (Washington University School of Medicine) for providing primary mouse skin fibroblasts. This study was supported by a Molecular Imaging Center grant from the US National Institutes of Health to Washington University and The University of Texas MD Anderson Cancer Center (P50 CA94056, D.P.-W.), by a Washington University Molecular Imaging Center Pilot Research Project grant (Z.Y.), by a grant from the US National Institutes of Health (R01GM098535, Z.Y.) and by an Interdisciplinary Research Initiative grant from the Children’s Discovery Institute of Washington University (MC-II-2012-215, Z.Y. and D.P.-W.). The High-Throughput Screening Core at the Washington University School of Medicine is supported in part by the Siteman Cancer Center (Cancer Center Support grant P30 CA91842 from the US National Cancer Institute).
PY - 2014/8
Y1 - 2014/8
N2 - The nonsense-mediated mRNA decay (NMD) pathway selectively eliminates aberrant transcripts containing premature translation termination codons and regulates the levels of a number of physiological mRNAs. NMD modulates the clinical outcome of a variety of human diseases, including cancer and many genetic disorders, and may represent a target for therapeutic intervention. Here, we have developed a new multicolored bioluminescence-based reporter system that can specifically and effectively assay NMD in live human cells. Using this reporter system, we conducted a robust high-throughput small-molecule screen in human cells and, unpredictably, identified a group of cardiac glycosides, including ouabain and digoxin, as potent inhibitors of NMD. Cardiac glycoside-mediated effects on NMD are dependent on binding and inhibiting the sodium-potassium ATPase on the plasma membrane and subsequent elevation of intracellular calcium levels. Induction of calcium release from the endoplasmic reticulum also leads to inhibition of NMD. Thus, this study reveals intracellular calcium as a key regulator of NMD and has implications for exploiting NMD in the treatment of disease.
AB - The nonsense-mediated mRNA decay (NMD) pathway selectively eliminates aberrant transcripts containing premature translation termination codons and regulates the levels of a number of physiological mRNAs. NMD modulates the clinical outcome of a variety of human diseases, including cancer and many genetic disorders, and may represent a target for therapeutic intervention. Here, we have developed a new multicolored bioluminescence-based reporter system that can specifically and effectively assay NMD in live human cells. Using this reporter system, we conducted a robust high-throughput small-molecule screen in human cells and, unpredictably, identified a group of cardiac glycosides, including ouabain and digoxin, as potent inhibitors of NMD. Cardiac glycoside-mediated effects on NMD are dependent on binding and inhibiting the sodium-potassium ATPase on the plasma membrane and subsequent elevation of intracellular calcium levels. Induction of calcium release from the endoplasmic reticulum also leads to inhibition of NMD. Thus, this study reveals intracellular calcium as a key regulator of NMD and has implications for exploiting NMD in the treatment of disease.
UR - http://www.scopus.com/inward/record.url?scp=84905817975&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84905817975&partnerID=8YFLogxK
U2 - 10.1038/nm.3620
DO - 10.1038/nm.3620
M3 - Article
C2 - 25064126
AN - SCOPUS:84905817975
SN - 1078-8956
VL - 20
SP - 961
EP - 966
JO - Nature medicine
JF - Nature medicine
IS - 8
ER -