Transcriptional Repression by Maf1 in Yeast

酵母中 Maf1 的转录抑制

基本信息

批准号：
7894176
负责人：
IAN M WILLIS
金额：
$ 22.04万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-13 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7894176
关键词：
Address Affect Bacteria Behavior Biochemical Biochemical Genetics Biological Biological Testing Cancer Biology Cell Cycle Cell Nucleus Cell Survival Cell division Cell physiology Cells Classification Complex Cyclic AMP-Dependent Protein Kinases Cytoplasm DNA Polymerase I DNA-Directed RNA Polymerase Databases Event Genetic Genetic Screening Genetic Transcription Goals Homologous Gene Human In Vitro Knowledge Link Mammalian Cell Mammals Maps Mediator of activation protein Metabolic Methods Modeling Modification Mutation Nuclear Nuclear RNA Nucleotides Nutritional Oncogenes Pathway interactions Phosphopeptides Phosphotransferases Polymerase Population Process Property Protein Kinase Proteins RNA Polymerase III Recombinants Recycling Regulation Repression Research Ribosomes Roentgen Rays Role Saccharomyces cerevisiae Signal Pathway Signal Repression Signal Transduction Signaling Molecule Site Solutions Specific qualifier value Stress Structure Structure-Activity Relationship System TP53 gene Testing Transcription Repressor/Corepressor Transfer RNA Tumor Suppressor Proteins Yeasts analytical ultracentrifugation c-myc Genes cost fitness gel electrophoresis gene repression genome-wide in vivo innovation insight loss of function mutation mutant novel protein folding protein structure public health relevance research study response three dimensional structure tumor

项目摘要

DESCRIPTION (provided by applicant): The synthesis of rRNAs and tRNAs is intimately linked with cell division through the function of tumor suppressors (e.g. p53 and Rb) and oncogenes (e.g. c-myc). The transforming forms of these proteins directly target and up-regulate transcription by RNA polymerases (pols) I and III along with the expression of important cell cycle regulators. The Maf1 protein is a unique regulator of transcription whose properties in human cells are similar to p53 and Rb. Human Maf1 is a candidate tumor suppressor that negatively regulates transcription by all three nuclear RNA polymerases. How Maf1 achieves its effects on transcription is not understood and is complicated by the fact that the protein does not contain any motifs of known function. In S. cerevisiae, Maf1 is essential for repressing transcription by RNA polymerase III and functions to integrate the responses from multiple nutritional and stress signaling pathways that coordinately regulate ribosome and tRNA synthesis. These pathways, like Maf1 itself, are not well defined. The universal requirement for Maf1 in pol III transcriptional repression in yeast provides an extraordinarily valuable model for understanding the pathways and mechanisms regulating ribosome and tRNA synthesis and the biochemical function of this novel signal integrator. Accordingly, the long-term goal of this research is to understand Maf1 structure/function relationships, its interactions with regulatory targets and its interactions with upstream signaling molecules that operate via posttranslational mechanisms to affect Maf1-dependent transcriptional repression. These aims will be achieved through biochemical studies in well-defined in vitro systems and through the application of an innovative method for phosphopeptide identification. In addition, powerful genome-wide and systematic genetic and biochemical approaches will be used to identify protein kinases and other molecules involved in signaling repression by Maf1. Finally, our understanding of the function and regulation of Maf1 will be advanced by the determination its three-dimensional structure. PUBLIC HEALTH RELEVANCE: Maf1 is a potential tumor suppressor that negatively regulates transcription by all three nuclear RNA polymerase in mammalian cells. In yeast, Maf1 is an essential mediator of transcriptional repression by RNA polymerase III and integrates the responses from multiple nutritional and stress signaling pathways that coordinately regulate ribosome and tRNA synthesis. Our genetic, biochemical and structural studies on on Maf1 will enhance understanding of fundamental cellular processes that are likely to impact cancer biology.

描述（由申请人提供）：RRNA和TRNA的合成通过肿瘤抑制器（例如p53和Rb）和肿瘤基因（例如C-MYC）的功能与细胞分裂密切相关。这些蛋白质的转化形式直接靶向RNA聚合酶（POLS）I和III以及重要细胞周期调节剂的表达。 MAF1蛋白是转录的独特调节剂，其在人类细胞中的特性与p53和rb相似。人MAF1是一种候选肿瘤抑制剂，对所有三种核RNA聚合酶进行负调节。 MAF1如何实现其对转录的影响尚不理解，并且由于蛋白质不包含已知功能的任何基序以下事实变得复杂。在酿酒酵母中，MAF1对于抑制RNA聚合酶III的转录至关重要，并功能可以整合来自多个营养和应力信号通路的反应，这些响应是协调调节核糖体和tRNA合成的。像MAF1本身一样，这些途径的定义不当。酵母中POL III转录抑制中MAF1的普遍要求为理解调节核糖体和TRNA合成的途径和机制提供了一个非常有价值的模型，以及该新型信号积分器的生化功能。因此，这项研究的长期目标是了解MAF1结构/功能关系，其与调节目标的相互作用及其与上游信号分子的相互作用，这些信号分子通过翻译后机制运行以影响MAF1依赖性转录抑制。这些目标将通过在体外系统明确的生化研究以及通过应用创新方法来实现磷酸肽鉴定来实现。此外，将使用强大的全基因组和系统的遗传和生化方法来鉴定MAF1信号抑制所涉及的蛋白激酶和其他分子。最后，我们对MAF1功能和调节的理解将通过确定其三维结构来提出。公共卫生相关性：MAF1是一种潜在的肿瘤抑制剂，对哺乳动物细胞中所有三种核RNA聚合酶的转录负调节。在酵母中，MAF1是RNA聚合酶III转录抑制的必不可少的介体，并整合了从多种营养和应力信号传导途径中的反应，这些反应协同调节了核糖体和tRNA合成。我们对MAF1的遗传，生化和结构研究将增强对可能影响癌症生物学的基本细胞过程的理解。