Analysis of Nonsense-Mediated mRNA Decay by Monitoring mRNA Half-Lives in Mammalian Cells

2010 ◽  
Vol 2010 (2) ◽  
pp. pdb.prot5386-pdb.prot5386 ◽  
Author(s):  
A. Yamashita ◽  
S. Ohno
2004 ◽  
Vol 24 (17) ◽  
pp. 7483-7490 ◽  
Author(s):  
Andrew Grimson ◽  
Sean O'Connor ◽  
Carrie Loushin Newman ◽  
Philip Anderson

ABSTRACT Eukaryotic messenger RNAs containing premature stop codons are selectively and rapidly degraded, a phenomenon termed nonsense-mediated mRNA decay (NMD). Previous studies with both Caenohabditis elegans and mammalian cells indicate that SMG-2/human UPF1, a central regulator of NMD, is phosphorylated in an SMG-1-dependent manner. We report here that smg-1, which is required for NMD in C. elegans, encodes a protein kinase of the phosphatidylinositol kinase superfamily of protein kinases. We identify null alleles of smg-1 and demonstrate that SMG-1 kinase activity is required in vivo for NMD and in vitro for SMG-2 phosphorylation. SMG-1 and SMG-2 coimmunoprecipitate from crude extracts, and this interaction is maintained in smg-3 and smg-4 mutants, both of which are required for SMG-2 phosphorylation in vivo and in vitro. SMG-2 is located diffusely through the cytoplasm, and its location is unaltered in mutants that disrupt the cycle of SMG-2 phosphorylation. We discuss the role of SMG-2 phosphorylation in NMD.


2013 ◽  
Vol 1829 (6-7) ◽  
pp. 612-623 ◽  
Author(s):  
Christoph Schweingruber ◽  
Simone C. Rufener ◽  
David Zünd ◽  
Akio Yamashita ◽  
Oliver Mühlemann

2009 ◽  
Vol 37 (6) ◽  
pp. 1287-1292 ◽  
Author(s):  
Lynne E. Maquat ◽  
Chenguang Gong

Nonsense-mediated mRNA decay and Staufen1-mediated mRNA decay are mechanistically related pathways that serve distinct purposes. In the present article, we give an overview of each pathway. We describe how a factor that is common to both pathways results in their competition. We also explain how competition between the two pathways contributes to the differentiation of C2C12 myoblasts to multinucleated myotubes.


Author(s):  
Jungyun Park ◽  
Seyoung Ahn ◽  
Aravinth K. Jayabalan ◽  
Takbum Ohn ◽  
Hyun Chul Koh ◽  
...  

2008 ◽  
Vol 36 (3) ◽  
pp. 514-516 ◽  
Author(s):  
Jikai Wen ◽  
Saverio Brogna

Translation and mRNA decay are coupled processes; the link is most obvious in the case of NMD (nonsense-mediated mRNA decay). NMD is a mechanism that drastically reduces the level of mRNA harbouring PTCs (premature translation termination codons). The defining event in NMD is premature translation termination and the key question is: what distinguishes premature from normal translation termination? Surprisingly, in mammalian cells, PTC recognition is linked to pre-mRNA splicing. Here, we review the current understanding in view of recent developments.


2018 ◽  
Author(s):  
Maximilian W. Popp ◽  
Lynne E. Maquat

AbstractDespite a long appreciation for the role of nonsense-mediated mRNA decay (NMD) in the destruction of faulty, disease-causing mRNAs, as well as its role in the maintenance of normal, endogenous transcript abundance, systematic unbiased methods for uncovering modifiers of NMD activity in mammalian cells remain scant. Here we present and validate a haploid genetic screening method for identifying proteins and processes that stimulate NMD activity involving a 3′-untranslated region exon-junction complex. This reporterbased screening method can be adapted for interrogating other pathways whose output can be measured by the intracellular production of fluorescent proteins.


2005 ◽  
Vol 12 (10) ◽  
pp. 893-901 ◽  
Author(s):  
Nao Hosoda ◽  
Yoon Ki Kim ◽  
Fabrice Lejeune ◽  
Lynne E Maquat

2014 ◽  
Vol 37 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Maximilian Wei-Lin Popp ◽  
Lynne E. Maquat

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