Premature termination codons in theDMDgene cause reduced local mRNA synthesis

Duchenne muscular dystrophy (DMD) is caused by mutations in theDMDgene leading to the presence of premature termination codons (PTC). Previous transcriptional studies have shown reduced DMD transcript levels in DMD patient and animal model muscles when PTC are present. Nonsense-mediated decay (NMD) has been suggested to be responsible for the observed reduction, but there is no experimental evidence supporting this claim. In this study, we aimed to investigate the mechanism responsible for the drop inDMDexpression levels in the presence of PTC. We observed that the inhibition of NMD does not normalizeDMDgene expression in DMD. Additionally, in situ hybridization showed that DMD messenger RNA primarily localizes in the nuclear compartment, confirming that a cytoplasmic mechanism like NMD indeed cannot be responsible for the observed reduction. Sequencing of nascent RNA to exploreDMDtranscription dynamics revealed a lower rate ofDMDtranscription in patient-derived myotubes compared to healthy controls, suggesting a transcriptional mechanism involved in reduced DMD transcript levels. Chromatin immunoprecipitation in muscle showed increased levels of the repressive histone mark H3K9me3 inmdxmice compared to wild-type mice, indicating a chromatin conformation less prone to transcription inmdxmice. In line with this finding, treatment with the histone deacetylase inhibitor givinostat caused a significant increase in DMD transcript expression inmdxmice. Overall, our findings show that transcription dynamics across theDMDlocus are affected by the presence of PTC, hinting at a possible epigenetic mechanism responsible for this process.

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Messenger RNA Transcripts of the Hepatocyte Nuclear Factor-1 Gene Containing Premature Termination Codons Are Subject to Nonsense-Mediated Decay

Diabetes ◽

10.2337/diabetes.53.2.500 ◽

2004 ◽

Vol 53 (2) ◽

pp. 500-504 ◽

Cited By ~ 41

Author(s):

L. W. Harries ◽

A. T. Hattersley ◽

S. Ellard

Keyword(s):

Messenger Rna ◽

Premature Termination ◽

Nuclear Factor ◽

Hepatocyte Nuclear Factor ◽

Nonsense Mediated Decay ◽

Rna Transcripts ◽

Premature Termination Codons ◽

Nuclear Factor 1

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mRNA surveillance: the perfect persist

Journal of Cell Science ◽

10.1242/jcs.115.15.3033 ◽

2002 ◽

Vol 115 (15) ◽

pp. 3033-3038 ◽

Cited By ~ 6

Author(s):

Eileen Wagner ◽

Jens Lykke-Andersen

Keyword(s):

Gene Expression ◽

Premature Termination ◽

Dominant Negative ◽

Nonsense Mediated Decay ◽

Premature Termination Codons ◽

Mrna Surveillance ◽

Multistep Process

In eukaryotes, an elaborate set of mechanisms has evolved to ensure that the multistep process of gene expression is accurately executed and adapted to cellular needs. The mRNA surveillance pathway works in this context by assessing the quality of mRNAs to ensure that they are suitable for translation. mRNA surveillance facilitates the detection and destruction of mRNAs that contain premature termination codons by a process called nonsense-mediated decay. Moreover, recent studies have shown that a distinct mRNA surveillance process, called nonstop decay, is responsible for depleting mRNAs that lack in-frame termination codons. mRNA surveillance thereby prevents the synthesis of truncated and otherwise aberrant proteins, which can have dominant-negative and other deleterious effects.

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The position of premature termination codons in the hepatocyte nuclear factor −1 beta gene determines susceptibility to nonsense-mediated decay

Human Genetics ◽

10.1007/s00439-005-0023-y ◽

2005 ◽

Vol 118 (2) ◽

pp. 214-224 ◽

Cited By ~ 36

Author(s):

L. W. Harries ◽

Coralie Bingham ◽

Christine Bellanne-Chantelot ◽

A. T. Hattersley ◽

Sian Ellard

Keyword(s):

Premature Termination ◽

Nuclear Factor ◽

Hepatocyte Nuclear Factor ◽

Nonsense Mediated Decay ◽

Premature Termination Codons ◽

Nuclear Factor 1 ◽

Beta Gene

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Genetic screens identify connections between ribosome recycling and nonsense mediated decay

10.1101/2021.08.03.454884 ◽

2021 ◽

Author(s):

Karole N D'Orazio ◽

Laura N. Lessen ◽

Anthony J. Veltri ◽

Zachary Neiman ◽

Miguel E. Pacheco ◽

...

Keyword(s):

Messenger Rna ◽

Premature Termination ◽

Stop Codon ◽

Premature Termination Codon ◽

Termination Codon ◽

Genetic Screens ◽

Regulatory Pathway ◽

Nonsense Mediated Decay ◽

Stop Codons ◽

Reported Data

The decay of messenger RNA with a premature termination codon (PTC) by nonsense mediated decay (NMD) is an important regulatory pathway for eukaryotes and an essential pathway in mammals. NMD is typically triggered by the ribosome terminating at a stop codon that is aberrantly distant from the poly-A tail. Here, we use a fluorescence screen to identify factors involved in NMD in S. cerevisiae . In addition to the known NMD factors, including the entire UPF family (UPF1, UPF2 and UPF3), as well as NMD4 and EBS1 , we identify factors known to function in post-termination recycling and characterize their contribution to NMD. We then use a series of modified reporter constructs that block both elongating and scanning ribosomes downstream of stop codons and demonstrate that a deficiency in recycling of 80S ribosomes or 40S subunits stabilizes NMD substrates. These observations in S. cerevisiae expand on recently reported data in mammals indicating that the 60S recycling factor ABCE1 is important for NMD (1,2) by showing that increased activities of both elongating and scanning ribosomes (80S or 40S) in the 3’UTR correlate with a loss of NMD.

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Identification of a Human Decapping Complex Associated with hUpf Proteins in Nonsense-Mediated Decay

Molecular and Cellular Biology ◽

10.1128/mcb.22.23.8114-8121.2002 ◽

2002 ◽

Vol 22 (23) ◽

pp. 8114-8121 ◽

Cited By ~ 230

Author(s):

Jens Lykke-Andersen

Keyword(s):

Saccharomyces Cerevisiae ◽

Premature Termination ◽

The Other ◽

Nonsense Mediated Decay ◽

Cell Extracts ◽

Rate Limiting Step ◽

Limiting Step ◽

Premature Termination Codons ◽

Rate Limiting ◽

Decapping Enzymes

ABSTRACT Decapping is a key step in general and regulated mRNA decay. In Saccharomyces cerevisiae it constitutes a rate-limiting step in the nonsense-mediated decay pathway that rids cells of mRNAs containing premature termination codons. Here two human decapping enzymes are identified, hDcp1a and hDcp2, as well as a homolog of hDcp1a, termed hDcp1b. Transiently expressed hDcp1a and hDcp2 proteins localize primarily to the cytoplasm and form a complex in human cell extracts. hDcp1a and hDcp2 copurify with decapping activity, an activity sensitive to mutation of critical hDcp residues. Importantly, coimmunoprecipitation assays demonstrate that hDcp1a and hDcp2 interact with the nonsense-mediated decay factor hUpf1, both in the presence and in the absence of the other hUpf proteins, hUpf2, hUpf3a, and hUpf3b. These data suggest that a human decapping complex may be recruited to mRNAs containing premature termination codons by the hUpf proteins.

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Two novel fibrillin-1 mutations resulting in premature termination codons but in different mutant transcript levels and clinical phenotypes

Human Mutation ◽

10.1002/humu.1380110112 ◽

1998 ◽

Vol 11 (S1) ◽

pp. S34-S37 ◽

Cited By ~ 11

Author(s):

Leena Karttunen ◽

Tarja Ukkonen ◽

Katariina Kainulainen ◽

Ann-Christine Syvänen ◽

Leena Peltonen

Keyword(s):

Premature Termination ◽

Transcript Levels ◽

Clinical Phenotypes ◽

Premature Termination Codons ◽

Mutant Transcript ◽

Fibrillin 1

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High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100166944 ◽

1996 ◽

Vol 54 ◽

pp. 896-897

Author(s):

G. W. Hacker ◽

I. Zehbe ◽

J. Hainfeld ◽

A.-H. Graf ◽

C. Hauser-Kronberger ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Messenger Rna ◽

High Performance ◽

Detection System ◽

Direct Methods ◽

Genetic Alterations ◽

Chain Reaction ◽

Protein Encoding ◽

Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

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