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 - Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
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
U2 - 10.1038/nm.3620
DO - 10.1038/nm.3620
M3 - Article
C2 - 25064126
AN - SCOPUS:84905817975
VL - 20
SP - 961
EP - 966
JO - Nature Medicine
JF - Nature Medicine
SN - 1078-8956
IS - 8
ER -