Regulation of RNA Polymerase II by the Ess1 Prolyl Isomerase

Ess1 脯氨酰异构酶对 RNA 聚合酶 II 的调节

• 批准号: 10158493
• 负责人: Steven D. HANES
• 金额: $ 32.4万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10158493
• 关键词: Address; Affect; Affinity; Antifungal Agents; Antineoplastic Agents; Area; Attenuated; Back; Binding; Binding Sites; Biochemical; Biological Assay; Biological Models; Biophysics; Candida albicans; Catalytic Domain; Cell Cycle Regulation; Cell Death; Cell Extracts; Cell physiology; Cells; ChIP-seq; Chemicals; Chromatin; Chromatin Remodeling Factor; Code; Complex; Consensus; Cryptococcus neoformans; Crystallization; Cyclic Amino Acids; DNA; Data; Development; Disease; Elongation Factor; Enzymes; Eukaryotic Cell; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genomic approach; Goals; Growth; HIV; Human; Immunocompromised Host; Immunosuppressive Agents; In Vitro; Instruction; Interferometry; Knowledge; Laboratories; Laboratory Study; Ligands; Link; Malignant Neoplasms; Maps; Mitotic Cell Cycle; Modeling; Modification; Molecular; Molecular Conformation; Mutation; Mycoses; Neurodegenerative Disorders; Organism; Orthologous Gene; Pathogenicity; Patients; Pattern; Peptides; Peptidylprolyl Isomerase; Phosphorylation; Physiological; Play; Positioning Attribute; Proline; Property; Proteins; RNA; RNA Polymerase II; RNA Processing; RNA Splicing; RNA polymerase II largest subunit; Reading; Regulation; Repetitive Sequence; Reporter; Research; Resolution; Roentgen Rays; Role; Saccharomyces cerevisiae; Series; Specific qualifier value; Structure; Substrate Domain; Substrate Specificity; Tertiary Protein Structure; Testing; Transcription Elongation; Transcriptional Regulation; Transplantation; Virulence; Work; Writing; Yeasts; analytical ultracentrifugation; cis trans isomerization; cis-trans-Isomerases; cofactor; experimental study; genome-wide; human disease; in vivo; inhibitor/antagonist; pathogenic fungus; protein protein interaction; recruit; sedimentation velocity; seryl-proline; stoichiometry; Address; Affect; Affinity; Antifungal Agents; Antineoplastic Agents; Area; Attenuated; Back; Binding; Binding Sites; Biochemical; Biological Assay; Biological Models; Biophysics; Candida albicans; Catalytic Domain; Cell Cycle Regulation; Cell Death; Cell Extracts; Cell physiology; Cells; ChIP-seq; Chemicals; Chromatin; Chromatin Remodeling Factor; Code; Complex; Consensus; Cryptococcus neoformans; Crystallization; Cyclic Amino Acids; DNA; Data; Development; Disease; Elongation Factor; Enzymes; Eukaryotic Cell; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genomic approach; Goals; Growth; HIV; Human; Immunocompromised Host; Immunosuppressive Agents; In Vitro; Instruction; Interferometry; Knowledge; Laboratories; Laboratory Study; Ligands; Link; Malignant Neoplasms; Maps; Mitotic Cell Cycle; Modeling; Modification; Molecular; Molecular Conformation; Mutation; Mycoses; Neurodegenerative Disorders; Organism; Orthologous Gene; Pathogenicity; Patients; Pattern; Peptides; Peptidylprolyl Isomerase; Phosphorylation; Physiological; Play; Positioning Attribute; Proline; Property; Proteins; RNA; RNA Polymerase II; RNA Processing; RNA Splicing; RNA polymerase II largest subunit; Reading; Regulation; Repetitive Sequence; Reporter; Research; Resolution; Roentgen Rays; Role; Saccharomyces cerevisiae; Series; Specific qualifier value; Structure; Substrate Domain; Substrate Specificity; Tertiary Protein Structure; Testing; Transcription Elongation; Transcriptional Regulation; Transplantation; Virulence; Work; Writing; Yeasts; analytical ultracentrifugation; cis trans isomerization; cis-trans-Isomerases; cofactor; experimental study; genome-wide; human disease; in vivo; inhibitor/antagonist; pathogenic fungus; protein protein interaction; recruit; sedimentation velocity; seryl-proline; stoichiometry

Project Summary The long-term goal is to understand the mechanism by which peptidyl-prolyl cis/trans isomerases (PPIases) function as molecular switches to regulate gene activity. PPIases catalyze the isomerization of the peptide bond that precedes the cyclic amino acid proline, causing conformational changes that facilitate the folding of newly-synthesized proteins and that regulate the activity of mature proteins by altering their activity or protein- protein interactions. PPIases are found in all organisms, and are best known as the targets of immunosuppressive drugs. However, their normal function in cells is poorly understood. We study a PPIase called Ess1, which is essential for growth in Saccharomyces cerevisiae. Ess1 and its human ortholog, Pin1 (which can substitute for Ess1 in yeast), are implicated in transcription regulation and mitotic cell cycle control. In the pathogenic fungi, Candida albicans and Cryptococcus neoformans, Ess1 is essential for virulence. In humans, misexpression of Pin1 may contribute to cancer and neurodegenerative disease. These findings make clear the importance of prolyl isomerization for cellular function. The goal of the proposed research is to understand the mechanism by which Ess1 recognizes and regulates RNA polymerase II (RNAPII) in eukaryotic cells. Work from this laboratory has shown that Ess1 binds and isomerizes peptidyl-prolyl bonds within the carboxy-terminal domain (CTD) of the large subunit of RNAPII. Ess1 acts by throwing a conformational “proline switch” thus plays an integral role in specifying the so-called “CTD code” that helps coordinate the recruitment of protein co-factors to the transcribing RNAPII complex. Ess1-induced conformational changes in the CTD are likely to be important for multiple steps in the transcription cycle. The goal of Aim1 is to understand how Ess1 “reads the CTD code,” a critical first step in its regulation of the RNAPII transcription cycle. To determine how Ess1 recognizes and binds the CTD a combination of structural, biochemical, biophysical and in vivo studies will be used. The results will be important for understanding how the mammalian ortholog (Pin1) recognizes RNAPII and a variety of other substrates. This information will also be useful for efforts to develop antifungal inhibitors (Ess1) or anti-cancer drugs (Pin1). The goal of Aim2 is to understand how Ess1 “writes the CTD code” by focusing on its role in transcription elongation. The working hypothesis is that Ess1 controls the recruitment and/or activity of elongation factors, and/or chromatin modifiers, thus regulating RNAPII elongation. This will be tested using biochemical and genomic approaches. These results will be important for two main reasons. First, they will help us understand how non-covalent proline switches (versus covalent modification such as phosphorylation – an area already heavily studied) regulate recruitment of RNAPII co-factors to the CTD. Second, they will help us understand how Ess1 regulates elongation, a mechanism that is increasingly recognized as critical in gene regulation. To our knowledge, this is the only proposal, and we are the only laboratory, studying the role of this key peptidyl-prolyl isomerase in transcription.

项目摘要 长期目标是了解肽基 - 丙酰基/trans的机制 异构酶（PPIASE）充当分子开关，以调节基因活性。 PPIASE催化循环氨基之前的胡椒键的异构化 酸脯氨酸，导致构象变化，促进新合成的折叠 蛋白质并通过改变其活性或蛋白质的活性来调节成熟蛋白的活性 蛋白质相互作用。 PPIAS在所有生物体中都发现，最著名的是靶标 免疫抑制药物。但是，它们在细胞中的正常功能知之甚少。 我们研究了一个名为ESS1的PPIASE，这对于酿酒酵母的生长至关重要。 ESS1及其人类直系同源物PIN1（可以代替酵母中的ESS1）与 转录调控和有丝分裂细胞周期控制。在致病真菌中，念珠菌 白色念珠菌和新生群的隐球菌对于病毒至关重要。在人类中 PIN1的表现可能有助于癌症和神经退行性疾病。这些 发现清楚地表明了丙酰异构化对细胞功能的重要性。 拟议的研究的目的是了解ESS1的机制 在真核细胞中识别和调节RNA聚合酶II（RNAPII）。从中工作 实验室表明，ESS1结合并异构化肽基 - 丙酰键 RNAPII大亚基的羧基末端结构域（CTD）。 ESS1通过投掷 因此，构象“脯氨酸开关”在指定所谓的“ CTD”中起着不可或缺的作用 代码”有助于协调蛋白质辅助因素募集到转录RNAPII 复杂的。 CTD中ESS1引起的构象变化可能对 转录周期中的多个步骤。 AIM1的目标是了解ESS1如何“读取CTD代码”，这是关键的第一步 在调节RNAPII转录周期中。确定ESS1如何认识和 结合CTD结构，生化，生物物理和体内研究的组合将 被使用。结果对于理解哺乳动物直系同源物（PIN1）将很重要 识别RNAPII和其他各种底物。此信息也将对 开发抗真菌抑制剂（ESS1）或抗癌药物（PIN1）的努力。 AIM2的目的是通过关注其“编写CTD代码”的方式来了解ESS1 在转录情节中的角色。工作假设是ESS1控制着招聘 和/或伸长因子的活性和/或染色质修饰剂，从而控制RNAPII 伸长。这将通过生化和基因组方法进行测试。这些结果 将很重要，这是两个主要原因。首先，他们将帮助我们了解非共价 脯氨酸开关（相对于共价修改，例如磷酸化 - 已经存在的区域 大量研究）调节RNAPII共同因素募集到CTD。第二，他们会帮助 我们了解ESS1如何调节伸长率，这种机制越来越被认可 对于基因调节至关重要。 据我们所知，这是唯一的建议，我们是研究角色的唯一实验室 在转录中的此关键肽基培养基异构酶。