TRANSCRIPTION ELONGATION FACTORS IN YEAST

基因表达的适当调节是正常所必需的。 生长和发育的发生。转录过程代表了 选择细胞将使用哪些遗传信息的第一步。 然而，选择合适的基因进行解码只是 开始了。一旦RNA聚合酶及其相关因子确定 正确的基因，转录过程必须确保准确和 有效延长，直到完成转录单位 解码了。也就是说，虽然对细胞哪些基因的准确选择 需要转录开始转录调控，高效 伸长决定遗传信息是否被转换 转化成对细胞有用的产品。在地球上有大量的基因 人类和其他真核细胞及其病毒 具体规定在延伸率的水平上。例如，这些 包括几个原癌基因(c-myc、c-myb、c-fos)、腺苷 脱氨酶，一组应激反应基因，以及HIV-1和HIV-2。 清楚地了解控制转录的机制 通过这些基因和其他基因的延伸是理解 正常发育以及癌症和艾滋病等疾病。 为了增加我们对成绩单控制的知识 伸长过程中，遗传和生化方法将是最 很强大。这种结合最容易在萌芽阶段进行 酵母，酿酒酵母。在那里有广泛的保护 蛋白质组分和转录的生化过程 真核生物。因此，通过对这种酵母菌的研究了解到的许多东西将 立即适用于哺乳动物细胞中的类似反应。这 该提案将解决有关监管 RNA聚合酶的延伸反应II.转录延伸 将对影响TFIIS的因素进行详细的分析，实验设计将 还确保新因素的识别和表征 除了参与延伸率控制的TFIIS。具体的 目标包括：(1)确定和表征功能替代物(S) 转录延伸TFIIS；(2)鉴定与之相互作用的蛋白质 TFIIS并确定其功能相关性；(3)鉴定氨基酸 酿酒酵母中TFIIS功能的重要残基；(4)鉴定 体内阻断酿酒酵母RNA聚合酶II伸长的位点 而且是在体外。

The proper regulation of gene expression is necessary in order for normal growth and development to occur. The transcription process represents the first step in selecting which genetic information the cell will use. However, the selection of the proper genes to decode is only the beginning. Once RNA polymerase and its associated factors identify the correct genes, the transcription process must insure accurate and efficient elongation until the complete transcription unit has been decoded. That is, while the accurate selection of which genes the cell needs to transcribe begins transcriptional regulation, efficient elongation determines whether or not the genetic information is converted into a product useful to the cell. There are a large number of genes in human, and other eukaryotic, cells and their viruses which are specifically regulated at the level of elongation. For example, these include several protooncogenes, (c-myc, c-myb, c-fos), adenosine deaminase, a collection of stress response genes, and HIV-1 and HIV-2. Clearly understanding the mechanisms which control transcription elongation through these and other genes is fundamental to understanding normal development as well as such diseases as cancer and AIDS. In order to increase our knowledge of the controls over the transcript elongation process, a genetic and biochemical approach would be most powerful. This combination is most easily carried out in the budding yeast, Saccharomyces cerevisiae. There is extensive conservation in the protein components and biochemical processes of transcription among eukaryotes. Thus, much of what is learned by studies of this yeast will be immediately applicable to similar reactions in mammalian cells. This proposal will address several questions about factors which regulate the elongation reaction of RNA polymerase II. While transcript elongation factor TFIIS will be analyzed in detail, the experimental design will also insure the identification and characterization of new factors in addition to TFIIS that participate in elongation control. The Specific Aims include: (1) Identify and characterize functional substitute(s) for transcript elongation TFIIS; (2) Identify proteins which interact with TFIIS and determine their functional relevance; (3) Identify amino acid residues important for TFIIS function in S. cerevisiae; (4) Identify sites which block elongation by S. cerevisiae RNA polymerase II in vivo and in vitro.