The Role of TFIIB in Transcription Activation

TFIIB 在转录激活中的作用

• 批准号: 6792170
• 负责人: LUIS A ESTRELLA-PEREZ
• 金额: $ 2.49万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-10 至 2005-09-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6792170
• 关键词: SDS polyacrylamide gel electrophoresis; circular dichroism; fungal genetics; gene expression; gene induction /repression; gene mutation; genetic transcription; homeostasis; microarray technology; northern blottings; polymerase chain reaction; predoctoral investigator; transcription factor; western blottings

DESCRIPTION (provided by applicant): Gene expression is regulated mainly at the level of transcription and transcription in turn, is regulated primarily at the level of initiation. Proper gene regulation is a critical event to maintain the homeostatic state of an organism. This is exemplified by diseases such as cancer, where improper gene regulation leads to uncontrolled cellular growth. Throughout the funding period of this proposal relevant information towards understanding the involvement of the general transcription factor TFIIB in the process of gene activation will be obtained. To comprehend how activator proteins work, a genetic strategy to identify physiological targets of the Pho4 and Adr1 transcription activators has been devised. Both proteins interact with TFIIB as part of their activation mechanism. The approach relies in the isolation of extragenic suppressors of the activation defect conferred by the deletion of these genes. To analyze the general role of TFIIB in transcription, a whole genome microarray analysis will be performed. The experiment will utilize TFIIB mutants at positions E62 and R78 that produce downstream shifts in transcription start site selection. To shed some light on the molecular basis of how do mutations at positions E62 and R78 of TFIIB affect gene expression, the conformational integrity of these mutants will be studied by means of circular dichroism (CD) analysis and by analyzing their susceptibility to proteases.

描述（由申请人提供）：基因表达主要在转录水平上调节，而转录又主要在起始水平上调节。 适当的基因调控是维持生物体内稳态的关键事件。例如癌症等疾病，基因调控不当会导致细胞生长失控。在本提案的整个资助期内，将获得了解一般转录因子TFIIB参与基因激活过程的相关信息。为了理解激活蛋白是如何工作的，设计了一种遗传策略来识别Pho4和Adr1转录激活因子的生理靶点。 这两种蛋白质与TFIIB相互作用作为其激活机制的一部分。 该方法依赖于这些基因的缺失所赋予的激活缺陷的基因外抑制子的分离。为了分析TFIIB在转录中的一般作用，将进行全基因组微阵列分析。 该实验将利用在转录起始位点选择中产生下游移位的位置E62和R78处的TFIIB突变体。为了阐明TFIIB的E62和R78位突变如何影响基因表达的分子基础，将通过圆二色性（CD）分析和分析其对蛋白酶的敏感性来研究这些突变体的构象完整性。