Regulation of Transcription Attenuation in the B. subtilis trp Operon

枯草芽孢杆菌 trp 操纵子中转录衰减的调节

• 批准号: 9118654
• 负责人: Paul Gollnick
• 金额: $ 49.8万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-01 至 1997-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9118654&HistoricalAwards=false
• 关键词: Regulation; Transcription; Attenuation; B.; subtilis

Transcription attenuation in the Bacillus subtilis trp operon is regulated by a trans-acting RNA binding protein, the product of the mtr locus. In the presence of tryptophan, Mtr recognizes, and presumably binds to, a site in the trp leader transcript. This binding influences the attenuator to form a transcription terminator. The mtr locus contains two structural genes, mtrA and mtrB; both are required to regulate the trp operon. The objective of this work is to determine how Mtr recognized its RNA target and how it influences the attenuator to cause transcription termination. Specific objectives are to: 1) purify Mtr and determine its subunit structure. 2) develop in vitro assays for Mtr function including RNA binding and transcription regulation. 3) characterize the RNA target site in the trp leader transcript and demonstrate that Mtr binds this site. Features of Mtr required for RNA-binding will also be studied. 4) determine the mechanism of tryptophan activation of Mtr. 5) determine if Mtr regulates other genes. The importance of RNA-binding proteins as gene regulators is clear. Sequence- specific RNA-protein interactions regulate a wide range of cellular processes. This work will provide information about how proteins recognize RNA and how RNA-binding proteins regulate gene expression.

枯草芽孢杆菌 trp 操纵子中的转录衰减受到反式作用 RNA 结合蛋白（mtr 基因座的产物）的调节。 在色氨酸存在的情况下，Mtr 识别并可能结合色氨酸前导转录本中的一个位点。 这种结合影响衰减子形成转录终止子。 mtr位点包含两个结构基因，mtrA和mtrB；两者都需要调节色氨酸操纵子。 这项工作的目的是确定 Mtr 如何识别其 RNA 靶标以及它如何影响衰减子导致转录终止。 具体目标是：1）纯化Mtr并确定其亚基结构。 2) 开发 Mtr 功能的体外测定，包括 RNA 结合和转录调控。 3) 表征 trp 前导转录本中的 RNA 靶位点，并证明 Mtr 结合该位点。 RNA 结合所需的 Mtr 特征也将被研究。 4)确定Mtr色氨酸激活机制。 5)确定Mtr是否调节其他基因。 RNA 结合蛋白作为基因调节剂的重要性是显而易见的。 序列特异性RNA-蛋白质相互作用调节广泛的细胞过程。 这项工作将提供有关蛋白质如何识别 RNA 以及 RNA 结合蛋白如何调节基因表达的信息。