CONTROL OF RIBOSOMAL GENE TRANSCRIPTION

核糖体基因转录的控制

• 批准号: 2174743
• 负责人: RONALD H REEDER
• 金额: $ 38.01万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2174743
• 关键词: RNA biosynthesis; X ray crystallography; biological signal transduction; cell growth regulation; chromatin; eukaryote; fungal genetics; gel mobility shift assay; genetic regulation; genetic transcription; laboratory rabbit; molecular cloning; molecular genetics; nucleosomes; protein structure; ribosomal RNA; tissue /cell culture; transcription factor

This project continues our long standing investigation into the molecular basis for regulating ribosomal RNA synthesis in eukaryotic cells. We have recently switched to yeast as our experimental organism and the proposed studies fall under four broad headings: 1) We will use a variety of genetic approaches to continue identification of genes and cloning of cDNAs for proteins that are part of the basal transcription initiation machinery utilized by RNA polymerase l (Poll). These genetic tools include a novel colony color assay we have developed in which yeast colonies change from dark red to light pink as Poll transcription is activated. 2) Pol I transcription activity is closely tied to the overall growth rate of the cell. We will use a combination of genetic and biochemical approaches to identify and study the signalling pathway which leads from the exterior of the cell to the Pol l machinery. 3) We have previously cloned a Poll transcription factor from the frog, Xenopus laevis, called xUBF. We will continue our studies of how this protein binds to DNA, to nucleosomes, and how it functions as a transcription factor. In collaboration with Dr. Barry Stoddard we will crystallize derivatives of xUBF and determine its 3-dimensional structure by X-ray crystallography. Both genetic and biochemical approaches will be used to identify the putative yeast homolog of xUBF. 4) Ongoing efforts to biochemically fractionate a yeast whole cell extract will be continued. Some transcription factors may be identified and cloned by this route. In addition, a well fractionated system is essential for detailed analysis of the molecular mechanisms that regulate Poll transcription. Ribosomal RNA comprises over half of the total RNA in a typical cell and it seems important to understand how synthesis of such a major cellular component is regulated. However, at present it is not clear whether this knowledge will have immediate clinical utility.

这个项目继续我们对 调节真核核糖体RNA合成的分子基础 细胞。 我们最近改用酵母作为我们的实验生物 拟议的研究属于四个广泛的标题： 1）我们将使用各种遗传方法继续 鉴定基因和CDNA的克隆是蛋白质的一部分 RNA使用的基底转录启动机制 聚合酶L（民意调查）。这些遗传工具包括一种新颖的殖民色彩 我们开发了酵母菌菌落从深红色变为的测定法 浅粉红色作为轮询转录被激活。 2）POL I转录活动与整体增长紧密相关 细胞的速率。我们将结合遗传和生化 识别和研究信号通路的方法 电池的外部到Pol L机械。 3）我们以前已经从青蛙中克隆了一个轮廓转录因子， Xenopus laevis，称为Xubf。我们将继续研究如何 蛋白质与DNA结合，与核小体以及它如何用作A 转录因子。与Barry Stoddard博士合作，我们将 结晶的XUBF衍生物并确定其3维 X射线晶体学结构。遗传和生化 方法将用于识别XUBF的假定酵母同系物。 4）正在进行的生物化学分馏酵母全细胞的努力 提取物将继续。可以确定一些转录因子 并用这条路线克隆。此外，分数良好的系统是 对于分子机制的详细分析至关重要的 调节民意调查转录。 核糖体RNA包含典型细胞中总RNA的一半以上，并且 了解如此主要的细胞的合成似乎很重要 组件受调节。但是，目前尚不清楚这是否 知识将具有立即的临床实用性。