Regulation of RNA polymerase ll by small RNAs

小RNA对RNA聚合酶II的调节

基本信息

  • 批准号:
    6802398
  • 负责人:
  • 金额:
    $ 29.41万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2003
  • 资助国家:
    美国
  • 起止时间:
    2003-09-19 至 2007-08-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): All cells respond to stress such as heat shock, UV irradiation, and viral infection, and do so in part by altering gene expression. A critical control point for regulating gene expression in eukaryotic cells is at the level of mRNA transcription by RNA polymerase II. In a break with paradigm, the preliminary data discussed here identify a natural RNA, mouse B2 RNA, which can bind to and inhibit transcription by RNA polymerase II. The proposed studies will investigate the role of B2 RNA as a transcriptional repressor in response to cellular stresses including heat shock, DNA damage, and viral infection. Completion of these studies will provide insight into how transcription is regulated under conditions of stress, which is vitally important in understanding normal cell growth as well as aberrations associated with diseases and deleterious environmental conditions. The specific aims of the proposal are the following: 1) To study the function of B2 RNA as a negative regulator of the transcriptional response to heat shock, DNA damage, and viral infection in mouse cells. 2) To determine the molecular mechanism by which B2 RNA inhibits transcription by RNA polymerase II. 3) To map the regions of B2 RNA and RNA polymerase II that interact, and in doing so develop a small nucleic acid inhibitor of the transcriptional response to stress. 4) To characterize human RNAs that bind to RNA polymerase II and regulate transcription in response to heat shock, DNA damage, and viral infection. The specific aims utilize both in vitro experiments and cell-based assays. The in vitro experiments employ a purified RNA polymerase II transcription system as well as nuclear extracts prepared from stressed mouse cells. The cell-based experiments utilize techniques to decrease levels of regulatory RNAs in cells such as antisense, ribozyme, and RNAi, as well as techniques to monitor RNA polymerase II activity such as nuclear run-on assays and chromatin immunoprecipitations. The proposed experiments have the potential to show that natural small noncoding RNAs can regulate transcription by targeting RNA polymerase II, to experimentally demonstrate a function for the SINE encoded B2 RNA, and to propose a mechanism by which general mRNA transcription is repressed in response to cellular stress.
描述(由申请人提供):所有细胞都会对热休克、紫外线照射和病毒感染等应激做出反应,并且部分是通过改变基因表达来实现的。真核细胞中调节基因表达的关键控制点是 RNA 聚合酶 II 的 mRNA 转录水平。打破范式,这里讨论的初步数据确定了一种天然 RNA,即小鼠 B2 RNA,它可以与 RNA 聚合酶 II 结合并抑制转录。拟议的研究将调查 B2 RNA 作为转录抑制因子在响应细胞应激(包括热休克、DNA 损伤和病毒感染)中的作用。这些研究的完成将深入了解应激条件下转录的调节方式,这对于理解正常细胞生长以及与疾病和有害环境条件相关的畸变至关重要。 该提案的具体目标如下: 1) 研究 B2 RNA 作为小鼠细胞热休克、DNA 损伤和病毒感染转录反应负调节因子的功能。 2) 确定B2 RNA抑制RNA聚合酶II转录的分子机制。 3) 绘制 B2 RNA 和 RNA 聚合酶 II 相互作用的区域图,并在此过程中开发出一种针对应激转录反应的小型核酸抑制剂。 4) 表征与 RNA 聚合酶 II 结合并调节转录以响应热休克、DNA 损伤和病毒感染的人类 RNA。 具体目标利用体外实验和基于细胞的测定。体外实验采用纯化的 RNA 聚合酶 II 转录系统以及从应激小鼠细胞中制备的核提取物。基于细胞的实验利用反义、核酶和 RNAi 等降低细胞中调节 RNA 水平的技术,以及监测 RNA 聚合酶 II 活性的技术,如核连续分析和染色质免疫沉淀。所提出的实验有可能表明天然小非编码RNA可以通过靶向RNA聚合酶II来调节转录,通过实验证明SINE编码的B2 RNA的功能,并提出一种响应细胞应激而抑制一般mRNA转录的机制。

项目成果

期刊论文数量(0)
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James Goodrich其他文献

James Goodrich的其他文献

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{{ truncateString('James Goodrich', 18)}}的其他基金

Do NFATc2 and cJun Cooperate to Activate Transcription in Breast Cancer Cells?
NFATc2 和 cJun 是否协同激活乳腺癌细胞的转录?
  • 批准号:
    8565637
  • 财政年份:
    2013
  • 资助金额:
    $ 29.41万
  • 项目类别:
Do NFATc2 and cJun Cooperate to Activate Transcription in Breast Cancer Cells?
NFATc2 和 cJun 是否协同激活乳腺癌细胞的转录?
  • 批准号:
    8706870
  • 财政年份:
    2013
  • 资助金额:
    $ 29.41万
  • 项目类别:
Do the B2 and Alu ncRNAs Repress Host Cell mRNA Transcription During HSV-1 Infect
B2 和 Alu ncRNA 在 HSV-1 感染期间抑制宿主细胞 mRNA 转录吗
  • 批准号:
    8503359
  • 财政年份:
    2012
  • 资助金额:
    $ 29.41万
  • 项目类别:
Do the B2 and Alu ncRNAs Repress Host Cell mRNA Transcription During HSV-1 Infect
B2 和 Alu ncRNA 在 HSV-1 感染期间抑制宿主细胞 mRNA 转录吗
  • 批准号:
    8600239
  • 财政年份:
    2012
  • 资助金额:
    $ 29.41万
  • 项目类别:
Regulation of Transcription at the Human IL-2 Promoter
人 IL-2 启动子的转录调控
  • 批准号:
    7936648
  • 财政年份:
    2009
  • 资助金额:
    $ 29.41万
  • 项目类别:
Regulation of Eukaryotic Transcription:Chromatin to mRNA
真核转录的调控:染色质到 mRNA
  • 批准号:
    7058654
  • 财政年份:
    2006
  • 资助金额:
    $ 29.41万
  • 项目类别:
Regulation of RNA Polymerase II by Small RNAs
小 RNA 对 RNA 聚合酶 II 的调节
  • 批准号:
    7486857
  • 财政年份:
    2003
  • 资助金额:
    $ 29.41万
  • 项目类别:
Regulation of RNA polymerase ll by small RNAs
小RNA对RNA聚合酶II的调节
  • 批准号:
    6942999
  • 财政年份:
    2003
  • 资助金额:
    $ 29.41万
  • 项目类别:
Regulation of RNA Metabolism by the RNA-dependent RNA Polymerase Activity of Mammalian RNA polymerase II
哺乳动物 RNA 聚合酶 II 的 RNA 依赖性 RNA 聚合酶活性对 RNA 代谢的调节
  • 批准号:
    9335918
  • 财政年份:
    2003
  • 资助金额:
    $ 29.41万
  • 项目类别:
Regulation of RNA Polymerase II by Non-coding RNAs
非编码 RNA 对 RNA 聚合酶 II 的调节
  • 批准号:
    8568673
  • 财政年份:
    2003
  • 资助金额:
    $ 29.41万
  • 项目类别:

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Dissecting the regulatory role of a eukaryotic transcription factor in RNA-templated transcription catalyzed by DNA-directed RNA polymerase II
剖析真核转录因子在 DNA 指导的 RNA 聚合酶 II 催化的 RNA 模板转录中的调节作用
  • 批准号:
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  • 财政年份:
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