Elucidating the essential role of the evolutionarily conserved IME4 mRNA methyltransferase in metazoan development

阐明进化上保守的 IME4 mRNA 甲基转移酶在后生动物发育中的重要作用

• 批准号: 8812310
• 负责人: CINTIA Fabiana HONGAY
• 金额: $ 34.86万
• 依托单位: CLARKSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8812310
• 关键词: Ablation; Academic Research Enhancement Awards; Adenosine; Adult; Affect; Animal Model; Animals; Antibodies; Binding; Biochemical; Biochemical Genetics; Bioinformatics; Biological; Brain; Categories; Cells; Cellular biology; Co-Immunoprecipitations; Complex; Critical Thinking; Data; Defect; Development; Developmental Biology; Dissection; Distant; Doctor of Philosophy; Drosophila genus; Embryo; Embryonic Development; Enzymes; Essential Genes; Evolution; Foundations; Gene-Modified; Genes; Genetic; Genetic Code; Germ Cells; Goals; Gonadal structure; Human; Immunoprecipitation; In Situ; Institution; Intestines; Invertebrates; Investigation; Life; Life Cycle Stages; Link; Mammals; Maps; Mass Spectrum Analysis; Meiosis; Mentors; Messenger RNA; Methylation; Methyltransferase; Microscopic; Microscopy; Modification; Molecular Biology; Mus; Oligonucleotides; Organ; Organism; Ovarian; Ovary; Parents; Pattern; Phase; Proteins; Publications; Publishing; Pupa; RNA; RNA Binding; RNA Splicing; RNA methylation; RNA-Protein Interaction; Reproduction; Research; Research Proposals; Research Technics; Role; Saccharomyces cerevisiae; Saccharomycetales; Sampling; Signal Transduction; Spliceosomes; Staging; Structure; Students; System; Techniques; Technology; Testing; Testis; Training; Transcript; United States National Institutes of Health; Universities; Vertebrates; Whole Organism; Work; Yeasts; Zebrafish; asexual; base; cohort; design; experience; graduate student; in vivo; knock-down; methylome; mutant; public health relevance; research study; response; skills; undergraduate student; zebrafish development

 DESCRIPTION (provided by applicant): Project Summary The proposal entitled 'Elucidating the essential role of the evolutionarily conserved IME4 mRNA methyltransferase in metazoan development' is in response to PAR 12-006 Academic Research Enhancement Award (Parent R15). The objective of this project is to elucidate the mechanism by which IME4 affects metazoan development, generating data for rational design to investigate its function in mammalian systems in the context of evolutionary conservation. IME4 (Inducer of Meiosis #4, first identified in budding yeast), is an N6methyltransferase of adenosine residues in messenger RNA without altering the genetic code, i.e., it is a non-editing RNA modification. IME4 is evolutionarily conserved from budding yeast to human beings, yet its biological role remains elusive. Recent publications have mapped the "mRNA methylome" in mice and human cells showing overlapping categories of this enzyme's putative targets, highlighting its evolutionarily conserved role. However, in vivo whole-organism studies of the biological role of this enzyme are lacking. My previous work in S. cerevisiae showed that this enzyme is key in budding yeast's decision to become a "gamete". It appears that IME4's role in yeast is exclusively during meiosis and it is not an essential gene for asexual life. However, in higher organisms where it has been studied, IME4 is an essential gene. In D.melanogaster, our published and preliminary data show it is required for embryo and larval development with important roles later in adult reproduction. Given its high degree of evolutionary conservation, we hypothesize that IME4 is required for essential functions in higher vertebrates, including humans. To test this hypothesis, we will employ biochemical (protein interactions), genetic, and cell biology (microscopy) approaches in D.melanogaster and D. rerio, two evolutionarily distant metazoan species, an invertebrate and a vertebrate respectively, predicting a high degree of functional conservation common to both that will be likely conserved in mammals. Approaches and techniques will provide excellent venues to train students in developing critical thinking skills and gain first-hand experience in the design and technology involved in biological experimentation. This project will generate important data that will impact the field of RNA modifications in developmental biology.

 描述（由申请人提供）： 项目摘要题为“阐明进化上保守的IME 4 mRNA甲基转移酶在后生动物发育中的重要作用”的提案是对PAR 12-006学术研究增强奖（Parent R15）的回应。该项目的目标是阐明IME 4影响后生动物发育的机制，为合理设计生成数据，以在进化保护的背景下研究其在哺乳动物系统中的功能。IME 4（减数分裂诱导剂#4，首先在芽殖酵母中鉴定）是信使RNA中腺苷残基的N6甲基转移酶，而不改变遗传密码，即，它是一种非编辑RNA修饰。从芽殖酵母到人类，IME 4在进化上是保守的，但它的生物学作用仍然难以捉摸。最近的出版物已经绘制了小鼠和人类细胞中的“mRNA甲基化组”，显示了这种酶的假定靶点的重叠类别，突出了其进化上保守的作用。然而，缺乏这种酶的生物学作用的体内全生物体研究。我以前在S的工作。酿酒酵母的研究表明，这种酶是芽殖酵母决定成为“配子”的关键。看来IME 4在酵母中的作用仅在减数分裂期间，并且它不是酵母中的必需基因。 无性生活然而，在已经研究的高等生物中，IME 4是一个必需基因。在D.melanogaster中，我们发表的和初步的数据表明，它是胚胎和幼虫发育所必需的，在成年生殖后期起着重要作用。鉴于其高度的进化保守性，我们假设IME 4是高等脊椎动物（包括人类）必需的基本功能。为了验证这一假设，我们将采用生物化学（蛋白质相互作用），遗传学和细胞生物学（显微镜）的方法在D.melanogaster和D. rerio，两个进化上遥远的后生动物物种，分别是无脊椎动物和脊椎动物，预测两者共同的高度功能保护可能在哺乳动物中被保护。方法和技术将为训练学生发展批判性思维技能并获得生物实验所涉及的设计和技术的第一手经验提供绝佳的场所。该项目将产生影响发育生物学中RNA修饰领域的重要数据。