Large scale investigation in yeast to identify novel gene(s) involved in mRNA non-stop decay pathway

Nonstop decay is an mRNA degradation pathway involved in identifying and eliminating nonstop transcripts. Compared to other RNA degradation pathways, very little is known about the NSD mechanism. To identify novel genes involved in NSD, we performed a large-scale analysis, and identified 68 gene candidates. From these results we picked three helicases, NAM7, ECM32, and SKI2 to further investigate. Spot test and colony count assay confirmed the role of selected candidates in NSD. The abundance of the nonstop mRNA was then evaluated using qRT-PCR method, and it was confirmed that the deletions of the selected candidates had no significant effect on nonstop gene at the transcriptional level when compared to the wildtype strain. Negative genetic interaction revealed association between candidate genes and translational regulation genes. The results of this study confirm the role of candidates in NSD but further research to characterize the genes.