THYMIDYLATE SYNTHASE GENE EXPRESSION

胸苷酸合酶基因表达

• 批准号: 2175473
• 负责人: LEE F JOHNSON
• 金额: $ 20.86万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-09-29 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2175473
• 关键词: THYMIDYLATE; SYNTHASE; GENE; EXPRESSION

Thymidylate synthase (TS) Is the enzyme that catalyzes the de novo biosynthesis of thymidylic acid. The enzyme Is essential in proliferating cells and is an important target enzyme In cancer chemotherapy. The overall goal of this project Is to understand the biochemical mechanisms for controlling the expression of the mouse TS gene In growth-stimulated cells. The studies In this proposal will focus on the Identification of the regulatory elements and trans-acting factors and explore In more detail the complex Interactions that are Important for proper regulation of this Important gene. Three specific aims are described. The first aim is to determine the locations of regulatory sequences that are upstream of the essential promoter elements. The trans-acting factors that Interact with the regulatory sequences will be studied. Qualitative or quantitative changes In these factors will be measured M cells progress from G1 through S phase. The possible relationship between these factors and the E2F transcription factor, or the RB tumor suppressor protein will be Investigated. If novel trans-acting factors are Identified, they will be characterized In detail. The second aim is to determine why Introns are necessary for proper regulation of TS minigenes. The ability of Introns from genes that are not cell cycle-regulated to restore proper regulation to an intronless TS minigene will be examined. The redundant (and perhaps autonomous) regulatory sequences that are located within Intron 2 will be Identified, and the mechanism by which they bring about growth- regulated expression will be examined. The third aim is to study in more detail the bidirectional nature of the mouse TS promoter. A preliminary characterization of the opposite direction promoter and the transcripts It may encode will be performed. The regulation of the opposite direction gene will be studied in growth-stimulated cells to determine if It is controlled In the same manner as the TS gene. If the 5' flanking sequences are found to block bidirectional transcription, the mechanism responsible for this inhibition will be analyzed. The proposed studies will increase our understanding of the mechanisms by which- cells regulate the progression from G1 to S phase. They will also provide additional-insight into the mechanisms by 'which sequences in the 5' flanking region of the gene as well as Introns affect gene expression.

胸苷酸合酶(TS)是催化从头合成的酶 胸腺酸的生物合成。这种酶在人体内是必不可少的 增殖细胞，是癌症中的重要靶酶 化疗。本项目的总体目标是了解 小鼠TS基因表达调控的生化机制 在生长刺激细胞中的基因。这项提案中的研究将集中在 论调控要素与交易行为的认定 因素，并更详细地探索复杂的交互作用 对这一重要基因的适当调控非常重要。三个具体的 对目标进行了描述。第一个目标是确定 基本启动子上游的调控序列 元素。与监管机构相互作用的交易因素 将对序列进行研究。这些变化的质或量的变化 测量因子将M细胞从G1期进展到S期。这个 这些因子与E2F转录的可能关系 因子，或Rb肿瘤抑制蛋白将被研究。如果 新的反式作用因子被识别出来，它们将在 细节。第二个目标是确定为什么内含子是必需的 对TS迷你菌的适当调节。基因内含子的能力 不受细胞周期调节的细胞，以恢复对 无内含子TS微型基因将被检测。多余的(也许还有 自治)位于内含子2内的调控序列将 以及它们带来增长的机制- 将检查受调控的表达。第三个目标是在更多的地方学习 详细说明小鼠TS启动子的双向性质。初赛 反向启动子和转录本的特性 它可以编码将被执行。对立面的规范 将在生长刺激细胞中研究方向基因以确定 如果它以与TS基因相同的方式控制。如果5英尺 侧翼序列被发现阻止双向转录， 对这种抑制的机理进行了分析。建议数 研究将增加我们对以下机制的了解： 细胞调控从G1期到S期的进程。他们还将 通过以下方式提供对机制的更多洞察： 基因的5‘侧翼区和内含子影响基因 表情。