Kal A J, van Zonneveld A J, Benes V, van den Berg M, Koerkamp M G, Albermann K, Strack N, Ruijter J M, Richter A, Dujon B, Ansorge W, Tabak H F
Department of Biochemistry, University of Amsterdam, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands.
Mol Biol Cell. 1999 Jun;10(6):1859-72. doi: 10.1091/mbc.10.6.1859.
We describe a genome-wide characterization of mRNA transcript levels in yeast grown on the fatty acid oleate, determined using Serial Analysis of Gene Expression (SAGE). Comparison of this SAGE library with that reported for glucose grown cells revealed the dramatic adaptive response of yeast to a change in carbon source. A major fraction (>20%) of the 15,000 mRNA molecules in a yeast cell comprised differentially expressed transcripts, which were derived from only 2% of the total number of approximately 6300 yeast genes. Most of the mRNAs that were differentially expressed code for enzymes or for other proteins participating in metabolism (e.g., metabolite transporters). In oleate-grown cells, this was exemplified by the huge increase of mRNAs encoding the peroxisomal beta-oxidation enzymes required for degradation of fatty acids. The data provide evidence for the existence of redox shuttles across organellar membranes that involve peroxisomal, cytoplasmic, and mitochondrial enzymes. We also analyzed the mRNA profile of a mutant strain with deletions of the PIP2 and OAF1 genes, encoding transcription factors required for induction of genes encoding peroxisomal proteins. Induction of genes under the immediate control of these factors was abolished; other genes were up-regulated, indicating an adaptive response to the changed metabolism imposed by the genetic impairment. We describe a statistical method for analysis of data obtained by SAGE.
我们描述了利用基因表达序列分析(SAGE)技术对在脂肪酸油酸酯上生长的酵母中mRNA转录水平进行的全基因组特征分析。将该SAGE文库与报道的在葡萄糖上生长的细胞的文库进行比较,揭示了酵母对碳源变化的显著适应性反应。酵母细胞中15,000个mRNA分子的很大一部分(>20%)包含差异表达的转录本,这些转录本仅来自约6300个酵母基因总数的2%。大多数差异表达的mRNA编码参与代谢的酶或其他蛋白质(如代谢物转运蛋白)。在油酸酯生长的细胞中,这表现为编码脂肪酸降解所需的过氧化物酶体β-氧化酶的mRNA大量增加。这些数据为跨细胞器膜存在氧化还原穿梭体提供了证据,这些穿梭体涉及过氧化物酶体、细胞质和线粒体酶。我们还分析了缺失PIP2和OAF1基因的突变菌株的mRNA谱,这两个基因编码诱导过氧化物酶体蛋白编码基因所需的转录因子。这些因子直接控制下的基因诱导被消除;其他基因上调,表明对基因损伤所导致的代谢变化的适应性反应。我们描述了一种用于分析SAGE获得的数据的统计方法。