Systematic approach for assessing whether undeletable chromosomal regions in Saccharomyces cerevisiae are required for cell viability
| Autor: | Satoshi Harashima, Keisuke Ekino, Naim Hassan, Farhana Easmin, Yu Sasano, Hisataka Taguchi |
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| Rok vydání: | 2020 |
| Předmět: |
Undeletable regions
Synthetic lethality lcsh:Biotechnology Saccharomyces cerevisiae lcsh:QR1-502 Biophysics Applied Microbiology and Biotechnology Genome lcsh:Microbiology 03 medical and health sciences 0302 clinical medicine Mini-chromosome loss lcsh:TP248.13-248.65 URA3 Gene 030304 developmental biology Suppressor mutation Genetics 0303 health sciences biology biology.organism_classification Phenotype Yeast Chromosomal region Original Article 030217 neurology & neurosurgery |
| Zdroj: | AMB Express AMB Express, Vol 10, Iss 1, Pp 1-17 (2020) |
| ISSN: | 2191-0855 |
| Popis: | Previously, we identified 49 undeletable chromosomal regions harboring only non-essential genes in the genome ofSaccharomyces cerevisiae. We proposed that there might be unknown synthetic lethal combinations of genes present in such undeletable regions of the genome. In this study, we chose four of the smallest undeletable chromosomal regions among the 49 and performed extensive further analyses to narrow down the gene-pairs responsible for lethality by replacing sub-regions in various combinations with a DNA module comprising theCgLEU2marker. Although the methodology was different from previous study, interestingly the results revealed that not only the sub-regions but also the entire region was replaceable. To solve the apparent discrepancy between previous and present results, we further conducted additional analysis including investigation of suppressor mutation and mini-chromosome loss assay through the construction of mini-chromosome harboring two particular chromosomal regions with marked withURA3marker by employing 5-FOA system. Based upon careful observation on the phenotype of colony formation on 5-FOA medium by spot test, we came to an important conclusion that particular chromosomal regions harboring only non-essential genes can be categorized into three classes, i.e., essential, non-essential and intrinsically essential. Intrinsically essential region is defined as appearance of papillae after mini-chromosome loss which implicates that the region is essential but compensatable against cell lethality. Our present study indicates that prudent and multiple approaches as performed in this study are needed to judge whether a particular chromosomal region of theS. cerevisiaegenome is essential, non-essential or intrinsically essential but compensatable. |
| Databáze: | OpenAIRE |
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