Nonsense codons can reduce the abundance of nuclear mRNA without affecting the abundance of pre-mRNA or the half-life of cytoplasmic mRNA.
Jiu Cheng,Lynne E. Maquat +1 more
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Experiments that placed TPI gene expression under the control of the human c-fos promoter, which can be transiently activated by the addition of serum to serum-deprived cells, verified that there was no detectable effect of a nonsense codon on the turnover of cytoplasmic mRNA.Abstract:
The abundance of the mRNA for human triosephosphate isomerase (TPI) is decreased to approximately 20% of normal by frameshift and nonsense mutations that cause translation to terminate at a nonsense codon within the first three-fourths of the reading frame. Results of previous studies inhibiting RNA synthesis with actinomycin D suggested that the decrease is not attributable to an increased rate of cytoplasmic mRNA decay. However, the step in TPI RNA metabolism that is altered was not defined, and the use of actinomycin D, in affecting all polymerase II-transcribed genes, could result in artifactual conclusions. In data presented here, the nonsense codon-mediated reduction in the level of TPI mRNA is shown to be characteristic of both nuclear and cytoplasmic fractions of the cell, indicating that the altered metabolic step is nucleus associated. Neither aberrancies in gene transcription nor aberrancies in RNA splicing appear to contribute to the reduction since there were no accompanying changes in the amount of nuclear run-on transcription, the level of any of the six introns in TPI pre-mRNA, or the size of processed mRNA in the nucleus. Deletion of all splice sites that reside downstream of a nonsense codon does not abrogate the reduction, indicating that the reduction takes place independently of the splicing of a downstream intron. Experiments that placed TPI gene expression under the control of the human c-fos promoter, which can be transiently activated by the addition of serum to serum-deprived cells, verified that there is no detectable effect of a nonsense codon on the turnover of cytoplasmic mRNA.read more
Citations
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Journal ArticleDOI
mRNA stability in mammalian cells.
TL;DR: This review concerns how cytoplasmic mRNA half-lives are regulated and how mRNA decay rates influence gene expression and techniques for measuring eukaryotic mRNA stability and for calculating decay constants.
Journal ArticleDOI
Nonsense-mediated mRNA decay: splicing, translation and mRNP dynamics
TL;DR: The acquisition and loss of mRNA-associated proteins accompanies the transition from the pioneer round to subsequent rounds of translation, and from translational competence to substrate for nonsense-mediated mRNA decay.
Journal ArticleDOI
Nonsense-Mediated mRNA Decay in Health and Disease
TL;DR: The current models of NMD that have been generated during the study of model organisms and mammalian cells are presented and the physiological burden of nonsense transcripts and the emerging view that NMD plays a broad and critical role in the regulation of gene expression are discussed.
Journal ArticleDOI
Quality control of eukaryotic mRNA: safeguarding cells from abnormal mRNA function
Olaf Isken,Lynne E. Maquat +1 more
TL;DR: An overview of three quality control mechanisms--nonsense-mediated RNA decay, nonstop mRNA decay, and no-go mRNA decay--is provided, which surveys mRNAs during translation and degrades those m RNAs that direct aberrant protein synthesis.
Journal ArticleDOI
Human Upf proteins target an mRNA for nonsense-mediated decay when bound downstream of a termination codon.
TL;DR: It is suggested that assembly of a dynamic hUpf complex initiates in the nucleus at mRNA exon-exon junctions and triggers NMD in the cytoplasm when recognized downstream of a translation termination site.
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