Cervelli Tiziana, Lodovichi Samuele, Bellè Francesca, Galli Alvaro
Yeast Genetics and Genomics Group, Laboratory of Functional Genetics and Genomics, Institute of Clinical Physiology CNR, Via Moruzzi 1, 56125 Pisa, Italy.
Microb Cell. 2020 May 18;7(7):162-174. doi: 10.15698/mic2020.07.721.
Technological advances are continuously revealing new genetic variants that are often difficult to interpret. As one of the most genetically tractable model organisms, yeast can have a central role in determining the consequences of human genetic variation. DNA repair gene mutations are associated with many types of cancers, therefore the evaluation of the functional impact of these mutations is crucial for risk assessment and for determining therapeutic strategies. Owing to the evolutionary conservation of DNA repair pathways between human cells and the yeast , several functional assays have been developed. Here, we describe assays for variants of human genes belonging to the major DNA repair pathways divided in functional assays for human genes with yeast orthologues and human genes lacking a yeast orthologue. Human genes with orthologues can be studied by introducing the correspondent human mutations directly in the yeast gene or expressing the human gene carrying the mutations; while the only possible approach for human genes without a yeast orthologue is the heterologous expression. The common principle of these approaches is that the mutated gene determines a phenotypic alteration that can vary according to the gene studied and the domain of the protein. Here, we show how the versatility of yeast can help in classifying cancer-associated variants.
技术进步不断揭示出往往难以解读的新基因变异。作为遗传上最易于处理的模式生物之一，酵母在确定人类基因变异的后果方面可发挥核心作用。DNA修复基因突变与多种癌症相关，因此评估这些突变的功能影响对于风险评估和确定治疗策略至关重要。由于人类细胞与酵母之间DNA修复途径的进化保守性，已开发出多种功能测定方法。在此，我们描述了针对属于主要DNA修复途径的人类基因变异的测定方法，这些测定方法分为针对具有酵母直系同源物的人类基因和缺乏酵母直系同源物的人类基因的功能测定。具有直系同源物的人类基因可以通过将相应的人类突变直接引入酵母基因或表达携带突变的人类基因来进行研究；而对于没有酵母直系同源物的人类基因，唯一可行的方法是异源表达。这些方法的共同原则是，突变基因会导致表型改变，这种改变可能因所研究的基因和蛋白质结构域而异。在此，我们展示了酵母的多功能性如何有助于对癌症相关变异进行分类。
