Nonsense-mediated mRNA decay (NMD) refers to a pathway used for purposes of surveillance towards creating a positive avenue through which to minimize errors that are likely to result from the expression of genes by enhancing the process of eliminating mRNA transcripts, which are believed to contain premature stop codons.
In higher eukaryotes, cells can recognize premature termination codon (PTC)-containing mRNAs through the translation of the mRNAs in a suboptimal environment. The expectation is that the cells will be in a position to determine the possibility of the subsequent ribosome, thereby triggering NMD. In S. cerevisiae, cells recognize PTC-containing mRNAs through a surveillance mechanism that helps in the identification of degraded transcripts that may contain these mRNAs. The surveillance mechanism is specially designed to evaluate different aspects associated with the mRNAs, including cellular homeostasis and cell cycle progression to when to trigger NMD.
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The types of events that occur within the cells that help in the generation of PTC-containing transcripts are molecular events. The events play a vital role in the assembly and subsequent activation of the NMD machinery.
Initially, the compound PTC124 was identified as a nonsense suppression agent. The identification of this agent was in a screen that can be described as high-throughput containing over 800,000 compounds while using a firefly luciferase-based readthrough reporter. The expectation was trying to determine whether the PTC suppression would result in any notable increment of firefly luciferase activity.
Aminoglycosides cause miscoding during translation by inhibiting bacterial protein synthesis. The outcome is that the aminoglycosides create a high possibility of a misreading of mRNA. That creates a significant impact concerning determining the general expectations in terms of ensuring that mRNAs are read accurately.
The main limitation to the widespread use of aminoglycosides is the fact that they tend to create a perception of reduced efficacy, seen explicitly in some circumstances. That means that their application is often limited to specific circumstances through which they can be considered as being proactive.
The results in Figure 2 and Figure 3a, b can be distinguished based on the elevated levels of creatine kinase, which can be seen in Figure 3a and b while the levels are considerably low in Figure 2. That creates a significant difference in the results of the dystrophin being produced by the PTC124-treated cultured in each of the samples.
The authors looked for broad, off-target effects of PTC124 by seeking to examine the cells within differentiated periods. Based on the results, the results were captured on a weekly basis for four weeks, which was essential to ensuring that the authors would be able to determine the broad, off-target effects of PTC124.
The rationale that defines thinking that authentic termination and premature termination codons might be selectively affected by PTC124 is that the dystrophin produced in the PTC124-treated cultured myotubes shows significant differentiation. That builds on the understanding that indeed, the codons are not affected similarly by PTC124; thus, leading to the differentiation in the dystrophin produced.
In seeking to argue that authentic termination codons are not the target of action of PTC124, the experiment performed is the proof-of-concept experiment. This experiment intends to highlight that indeed, the target of action of PTC124 is not, in any way, the target for authentic termination codons.
The potential mechanistic explanation for increased protein production after treatment with PTC124 is that there exists a major difference between premature and normal termination, which seeks to reflect on some of the experimental factors leading to increased protein production. The possibility of this mechanistic explanation is experimentally distinguished through the proof-of-concept experiment that can point to the main factors contributing to the increased protein production.
