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

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.

项目总结

项目成果

Sec61 channel subunit Sbh1/Sec61β promotes ER translocation of proteins with suboptimal targeting sequences and is fine-tuned by phosphorylation.

• DOI: 10.1016/j.jbc.2023.102895
• 发表时间: 2023-03
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Barbieri, Guido;Simon, Julien;Lupusella, Cristina R.;Pereira, Fabio;Elia, Francesco;Meyer, Hadar;Schuldiner, Maya;Hanes, Steven D.;Nguyen, Duy;Helms, Volkhard;Roemisch, Karin
• 通讯作者: Roemisch, Karin