Regulation of transcriptional initiation by the 19S proteasome subcomplex.

19S 蛋白酶体亚复合体对转录起始的调节。

• 批准号: 8497039
• 负责人: Sukesh R. Bhaumik
• 金额: $ 33.63万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2017-09-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8497039
• 关键词: 26S proteasome; ATP phosphohydrolase; Acetyltransferase; Address; Applications Grants; Biochemical; Cell Cycle; Cell physiology; Cells; Complex; Cyclin-Dependent Kinase Inhibitor; Cyclins; DNA Microarray Chip; Development; Disease; Eukaryota; Eukaryotic Cell; Funding; Galactose; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Histone H4; Human; Investigation; Life; Link; Malignant Neoplasms; Mediator of activation protein; Methodology; Modeling; Nucleosome Core Particle; Nucleosomes; Oncogene Proteins; Ornithine Decarboxylase; Outcomes Research; Play; Proteins; RNA Polymerase II; Recruitment Activity; Regulation; Research; Research Proposals; Resources; Role; Series; Structure; Study Section; Therapeutic Agents; Transcription Initiation; Transcriptional Regulation; United States National Institutes of Health; Yeasts; base; chromatin immunoprecipitation; genome-wide; in vivo; multicatalytic endopeptidase complex; particle; promoter; protein degradation; public health relevance; research study; transcription factor; 26S proteasome; ATP phosphohydrolase; Acetyltransferase; Address; Applications Grants; Biochemical; Cell Cycle; Cell physiology; Cells; Complex; Cyclin-Dependent Kinase Inhibitor; Cyclins; DNA Microarray Chip; Development; Disease; Eukaryota; Eukaryotic Cell; Funding; Galactose; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Histone H4; Human; Investigation; Life; Link; Malignant Neoplasms; Mediator of activation protein; Methodology; Modeling; Nucleosome Core Particle; Nucleosomes; Oncogene Proteins; Ornithine Decarboxylase; Outcomes Research; Play; Proteins; RNA Polymerase II; Recruitment Activity; Regulation; Research; Research Proposals; Resources; Role; Series; Structure; Study Section; Therapeutic Agents; Transcription Initiation; Transcriptional Regulation; United States National Institutes of Health; Yeasts; base; chromatin immunoprecipitation; genome-wide; in vivo; multicatalytic endopeptidase complex; particle; promoter; protein degradation; public health relevance; research study; transcription factor

DESCRIPTION (provided by applicant): The 26S proteasome complex is essential for targeted protein degradation. It is composed of a 20S core particle (CP) and a 19S regulatory particle (RP). The 20S CP confers the proteolytic activity to the proteasome. The 19S RP has six different ATPase components, and regulates the function of the 20S CP in an ATP-dependent manner. Intriguingly, several studies have implicated the non-proteolytic role of the 19S RP in transcriptional initiation or activation independently of the 20S CP. However, the detailed regulatory mechanisms of transcriptional initiation by the 19S RP remained largely unknown in living cells. Based on our current NIH R15 funding, we have demonstrated in yeast that the 19S RP, but not 20S CP, is recruited to the GAL1 UAS (upstream activating sequence), and establishes a specific interaction network of Gal4-SAGA- Mediator (SAGA, Spt-Ada-Gcn5-acetyltransferase) at the UAS by enhancing the interaction of activator Gal4 with coactivator SAGA, which promotes the formation of preinitiation complex (PIC) at the core promoter to initiate transcription. We subsequently found similar results at several other genes. Overall, we find that 19S RP promotes the interaction of activator with coactivator for facilitating the formation of PIC, and hence transcriptional initiation. However, it is not clearly understood how the 19S RP promotes the interaction of activator with coactivator. Further, the roles of different ATPases of the 19S RP in such regulation are yet unknown. Moreover, it is not known whether the 19S RP functions as a global regulator of transcriptional initiation. Answers to these important questions will significantly contribute to the regulatory roles and mechanisms of the 19S RP in transcriptional initiation. Therefore, in this NIH R15 renewal application, we propose to address these questions. We will specifically determine (i) the role of the 19S RP in global transcriptional initiation, and (ii) how the 19S RP or its ATPase activity promotes the activator-coativator interaction, and hence transcriptional initiation. We will address these specific research aims primarily using the ChIP (chromatin immunoprecipitation), mutational, transcriptional, biochemical, biophysical, and ChIP- seq methodologies. The outcomes of this research proposal will provide important information on the 19S RP regulation of eukaryotic transcriptional initiation, and hence will significantly contribute to our current understanding of eukaryotic gene regulation. Since the transcription machinery as well as the 19S RP is conserved throughout eukaryotes from yeast to humans, these results in yeast will provide important resources for research in humans. Thus, the results will assist in the development of transcription-based therapeutic agents, as a growing number of diseases including cancer are linked to aberrant transcriptional initiation and/or are characterized by altered patterns of gene expression.

描述（由申请人提供）：26S蛋白酶体复合物对于靶向蛋白质降解至关重要。它由20S核心粒子（CP）和19S调节粒子（RP）组成。 20S CP将蛋白水解活性赋予蛋白酶体。 19S RP具有六个不同的ATPase组件，并以ATP依赖性方式调节20S CP的功能。有趣的是，几项研究暗示了19S RP在转录启动或激活中独立于20S CP的非蛋白水解作用。但是，19S RP的转录启动的详细调节机制在活细胞中基本上尚不清楚。 Based on our current NIH R15 funding, we have demonstrated in yeast that the 19S RP, but not 20S CP, is recruited to the GAL1 UAS (upstream activating sequence), and establishes a specific interaction network of Gal4-SAGA- Mediator (SAGA, Spt-Ada-Gcn5-acetyltransferase) at the UAS by enhancing the interaction of activator Gal4 with coactivator SAGA, which促进在核心启动子处的前启动复合物（PIC）的形成，以启动转录。随后，我们在其他几个基因上发现了类似的结果。总体而言，我们发现19S RP促进了激活剂与共激活因子的相互作用，以促进PIC的形成，从而促进了转录启动。但是，尚未清楚地理解19S RP如何促进激活剂与共激活因子的相互作用。此外，19S RP在这种调节中的不同ATPases的作用尚不清楚。此外，尚不清楚19S RP是否充当转录启动的全球调节剂。这些重要问题的答案将大大有助于19S RP在转录启动中的调节作用和机制。因此，在此NIH R15更新应用中，我们建议解决这些问题。我们将明确确定（i）19S RP在全球转录启动中的作用，以及（ii）19S RP或其ATPase活性如何促进激活剂式摄取器的相互作用，从而如何促进转录启动。我们将主要使用CHIP（染色质免疫沉淀），突变，转录，生化，生物物理和芯片Seq方法来解决这些特定研究。该研究建议的结果将提供有关真核转录启动的19S RP调节的重要信息，因此将极大地有助于我们当前对 真核基因调节。由于从酵母到人类的整个真核生物中保存了转录机械以及19S RP，因此这些酵母中的这些结果将为人类研究提供重要的资源。因此，结果将有助于开发基于转录的治疗剂，因为越来越多的疾病在内，包括癌症的疾病与异常的转录起始有关，并且/或以基因表达的模式改变。