Juneau Kara, Palm Curtis, Miranda Molly, Davis Ronald W
Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA.
Proc Natl Acad Sci U S A. 2007 Jan 30;104(5):1522-7. doi: 10.1073/pnas.0610354104. Epub 2007 Jan 23.
Knowing gene structure is vital to understanding gene function, and accurate genome annotation is essential for understanding cellular function. To this end, we have developed a genome-wide assay for mapping introns in Saccharomyces cerevisiae. Using high-density tiling arrays, we compared wild-type yeast to a mutant deficient for intron degradation. Our method identified 76% of the known introns, confirmed 18 previously predicted introns, and revealed 9 formerly undiscovered introns. Furthermore, we discovered that all 13 meiosis-specific intronic yeast genes undergo regulated splicing, which provides posttranscriptional regulation of the genes involved in yeast cell differentiation. Moreover, we found that approximately 16% of intronic genes in yeast are incompletely spliced during exponential growth in rich medium, which suggests that meiosis is not the only biological process regulated by splicing. Our tiling-array assay provides a snapshot of the spliced transcriptome in yeast. This robust methodology can be used to explore environmentally distinct splicing responses and should be readily adaptable to the study of other organisms, including humans.
了解基因结构对于理解基因功能至关重要,而准确的基因组注释对于理解细胞功能必不可少。为此,我们开发了一种全基因组分析方法来绘制酿酒酵母中的内含子图谱。利用高密度平铺阵列,我们将野生型酵母与缺乏内含子降解能力的突变体进行了比较。我们的方法鉴定出了76%的已知内含子,证实了18个先前预测的内含子,并发现了9个以前未被发现的内含子。此外,我们发现所有13个减数分裂特异性内含子酵母基因都经历了调控剪接,这为酵母细胞分化所涉及的基因提供了转录后调控。此外,我们发现,在丰富培养基中指数生长期间,酵母中约16%的内含子基因剪接不完全,这表明减数分裂不是唯一受剪接调控的生物学过程。我们的平铺阵列分析提供了酵母中剪接转录组的快照。这种强大的方法可用于探索环境不同的剪接反应,并且应该很容易适用于包括人类在内的其他生物体的研究。