Mechanisms of Splicing Regulation During Environmental Stress in Yeast

环境胁迫下酵母剪接调控机制

• 批准号: 8122997
• 负责人: Jaclyn C. Greimann
• 金额: $ 5.13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8122997
• 关键词: 3' Untranslated Regions; Affect; Amino Acids; Amitrole; Biological Assay; Cells; Cellular Stress; Cessation of life; Collection; Complement; DNA Damage; DNA Replication Damage; Disease; Enzyme Gene; Enzymes; Essential Genes; Evolution; Exposure to; Family; Foundations; Future; Gene Expression; Gene Mutation; Genes; Genomics; Goals; Introns; Investigation; Lead; Lysine; Mammals; Messenger RNA; Methods; Molecular; Monitor; Mutation; Pathway interactions; Pattern; Phenotype; Phosphotransferases; Process; Proliferating Cell Nuclear Antigen; Protein Kinase; Proteins; RNA; RNA Splicing; RNA purification; Reaction; Regulation; Regulatory Pathway; Research Proposals; Retinitis Pigmentosa; Reverse Transcriptase Polymerase Chain Reaction; Ribosomal Proteins; Saccharomyces cerevisiae; Spliced Genes; Starvation; Stimulus; Stress; Tail; Temperature; Testing; Trans-Activators; Ubiquitin; Ubiquitination; Yeasts; attenuation; base; biological adaptation to stress; casein kinase II; extracellular; falls; follow-up; genetic element; genome wide association study; interest; mRNA Precursor; mutant; promoter; research study; response; small molecule; transcription termination

DESCRIPTION (provided by applicant): Less than 5% of Saccharomyces cerevisiae genes have introns and most yeast intron-containing genes have only one intron. This contrasts with many other species in which the majority of genes possess multiple introns. It is possible that yeast retained only introns with an important function throughout evolution. Interestingly, the yeast intron-containing genes fall into only a few classes such as ribosomal protein genes, genes involved in sporulation and ubiquitin-like conjugation enzymes (Grate and Ares, 2002), suggesting a possible function of splicing in these pathways. In support of this, ribosomal protein gene introns accumulate upon amino acid starvation, conditions which lead translational stress (Pleiss et al., 2007a). This observation is the foundation for future experiments aiming to understand the starvation splicing regulatory mechanism as well as additional environmental stresses. All cells must respond to multiple environmental stress conditions and an improper response can lead to cellular death or disease. The aim of this proposal is to determine if cells employ splicing regulation as an additional mechanism to deal with stress. ) Preliminary results show both cis- and trans-acting factors are involved in the starvation splicing regulatory pathway. The objective of Aim 1 is to follow up preliminary results showing ribosomal protein gene promoters and not the introns are essential for the starvation splicing response and to determine the sequences specifically regulating this response. There is also evidence that a protein kinase, casein kinase 2, is necessary for the splicing response. Aim 2 focuses on determining additional factors involved in the regulatory pathway and will employ a genome-wide screen. And finally, the goal of Aim 3 is to assess whether other intron-rich gene classes are regulated at the level of splicing in response to environmental stress by examining additional cellular stress conditions, such as DNA damage. RNA undergoes many different processing steps, in addition to splicing, throughout gene expression. When any of these processing pathways is incorrectly regulated in mammals, diseases arise such as Retinitis pigmentosa caused by mutation of proteins involved in splicing (Cooper et al., 2009). A greater understanding of splicing regulatory pathways will not only allow for a more detailed view of splicing regulation in yeast, but also the many mechanisms altering gene expression during environmental stress. PUBLIC HEALTH RELEVANCE: During gene expression, RNA serves as a transient message which undergoes many molecular changes, or processing reactions, which alter its stability including 5' cap formation, RNA splicing and addition of a polyadenine tail. Cells regulate each of these reactions in response to extracellular stimuli or environmental stress and disruption of many of these processing mechanisms leads to disease. The goal of this research proposal is to understand the mechanisms regulating RNA splicing in response to environmental stress.

描述（由申请人提供）：酿酒酵母基因中只有不到5％的含有内含子，大多数含酵母内含子内含子的基因只有一个内含子。这与大多数基因具有多个内含子的许多其他物种形成鲜明对比。酵母可能仅保留在整个演化过程中具有重要功能的内含子。有趣的是，含有酵母内含子含有的基因仅属于少数几类，例如核糖体蛋白基因，涉及孢子型和泛素样共轭酶的基因（Grete and Ares，2002），这表明在这些途径中旋转的可能功能。为了支持这一点，核糖体蛋白基因内含子在氨基酸饥饿后积累，导致转化应激的条件（Pleiss等，2007a）。该观察结果是未来实验的基础，旨在了解饥饿的剪接调节机制以及其他环境应力。所有细胞都必须对多种环境应力条件做出反应，并且反应不当会导致细胞死亡或疾病。该提案的目的是确定细胞是否采用剪接调节作为应对压力的附加机制。 ）初步结果表明，饥饿的调节途径均参与顺式和反式作用因子。目标1的目的是跟踪显示核糖体蛋白基因启动子而不是内含子的初步结果，而不是内含子对于饥饿的剪接响应至关重要，并确定明确调节该反应的序列。也有证据表明，蛋白激酶酪蛋白激酶2对于剪接反应是必需的。 AIM 2专注于确定监管途径中涉及的其他因素，并将采用全基因组筛查。最后，目标3的目的是评估是否通过检查其他细胞应激条件（例如DNA损伤）来评估其他富含富含器的基因类别在剪接水平上受到剪接水平的调节。在整个基因表达中，RNA除了剪接外，还遵循许多不同的处理步骤。当这些加工途径中的任何一个在哺乳动物中受到不正确的调节时，会引起疾病，例如由涉及剪接的蛋白质突变引起的视网膜炎色素炎（Cooper等，2009）。对剪接调节途径的更多了解不仅允许对酵母中的剪接调节的更详细的看法，而且还允许在环境压力期间改变基因表达的许多机制。 公共卫生相关性：在基因表达期间，RNA充当瞬态信息，它经历了许多分子变化或加工反应，这会改变其稳定性，包括5'cap形成，RNA剪接和添加多丹宁尾巴。细胞对细胞外刺激或环境压力的响应调节这些反应中的每一个，以及许多此类加工机制的破坏导致疾病。该研究建议的目的是了解调节RNA剪接的机制，以应对环境压力。