Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario M5S 3E1, Canada.
Genetics. 2013 Sep;195(1):275-87. doi: 10.1534/genetics.113.152918. Epub 2013 Jul 12.
Whole-genome sequencing, particularly in fungi, has progressed at a tremendous rate. More difficult, however, is experimental testing of the inferences about gene function that can be drawn from comparative sequence analysis alone. We present a genome-wide functional characterization of a sequenced but experimentally understudied budding yeast, Saccharomyces bayanus var. uvarum (henceforth referred to as S. bayanus), allowing us to map changes over the 20 million years that separate this organism from S. cerevisiae. We first created a suite of genetic tools to facilitate work in S. bayanus. Next, we measured the gene-expression response of S. bayanus to a diverse set of perturbations optimized using a computational approach to cover a diverse array of functionally relevant biological responses. The resulting data set reveals that gene-expression patterns are largely conserved, but significant changes may exist in regulatory networks such as carbohydrate utilization and meiosis. In addition to regulatory changes, our approach identified gene functions that have diverged. The functions of genes in core pathways are highly conserved, but we observed many changes in which genes are involved in osmotic stress, peroxisome biogenesis, and autophagy. A surprising number of genes specific to S. bayanus respond to oxidative stress, suggesting the organism may have evolved under different selection pressures than S. cerevisiae. This work expands the scope of genome-scale evolutionary studies from sequence-based analysis to rapid experimental characterization and could be adopted for functional mapping in any lineage of interest. Furthermore, our detailed characterization of S. bayanus provides a valuable resource for comparative functional genomics studies in yeast.
全基因组测序,特别是在真菌中,发展速度非常快。然而,更困难的是仅通过比较序列分析来实验测试关于基因功能的推断。我们对已测序但实验研究不足的出芽酵母 Saccharomyces bayanus var. uvarum(以下简称 S. bayanus)进行了全基因组功能特征描述,使我们能够绘制出该生物体与 S. cerevisiae 分离的 2000 万年的变化图谱。我们首先创建了一套遗传工具,以方便在 S. bayanus 中进行工作。接下来,我们使用一种计算方法来优化各种扰动,测量了 S. bayanus 对多样化的基因表达响应,以涵盖广泛的功能相关的生物学反应。由此产生的数据集表明,基因表达模式在很大程度上是保守的,但在调节网络中可能存在显著变化,如碳水化合物利用和减数分裂。除了调节变化,我们的方法还确定了已经分化的基因功能。核心途径中的基因功能高度保守,但我们观察到许多基因参与渗透胁迫、过氧化物酶体生物发生和自噬的变化。大量特定于 S. bayanus 的基因对氧化应激有反应,这表明该生物体可能经历了与 S. cerevisiae 不同的选择压力。这项工作将基因组规模的进化研究从基于序列的分析扩展到快速实验特征描述,并可用于任何感兴趣的谱系的功能映射。此外,我们对 S. bayanus 的详细描述为酵母中比较功能基因组学研究提供了有价值的资源。