The role of DOT1L methyltransferase in controlling the noncoding transcriptome

DOT1L 甲基转移酶在控制非编码转录组中的作用

基本信息

批准号：
10669669
负责人：
Alla Grishok
金额：
$ 33万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10669669
关键词：
Animals Antibodies Antisense RNA Antiviral Response Binding Binding Proteins Caenorhabditis elegans Cell Nucleus Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Code Complementary RNA Complex Congenital Abnormality Cytoplasm Data Development Developmental Gene Double-Stranded RNA Enhancers Enzymes Experimental Genetics Fishes Future Gene Expression Gene Expression Regulation Gene Silencing Generations Genes Genetic Genetic Transcription Genomic approach Genomics Goals Histone H2A Histone H3 Homologous Gene Human Immunoprecipitation Laboratories Link Lysine Malignant Neoplasms Mammalian Cell Mammals Mediator Messenger RNA Methylation Methyltransferase MicroRNAs Modeling Molecular Mutation Names Nematoda Nuclear RNA Nucleotides PHD Finger Pathogenesis Pathway interactions Phenotype Plants Process Production Proteins Publishing RNA RNA Binding RNA Interference RNA-Protein Interaction Research Role Small Interfering RNA Small RNA System Testing Titrations Transcriptional Activation Transcriptional Regulation Transgenes Untranslated RNA Variant Viral Virus Work Yeasts Zebrafish Zinc Fingers base deep sequencing genetic approach genome wide methylation global run on sequencing human disease insight interest leukemia loss of function mRNA Precursor malignant breast neoplasm mutant novel promoter protein complex recruit transcriptome

项目摘要

The broad goal of our research is to understand how gene expression is regulated during animal development. Our specific interest is in finding natural regulators of endogenous double-stranded RNA production and nuclear RNA interference. In this regard, we are focusing on the relationship between RNA interference and the histone H3 lysine 79 DOT1L methyltransferase complex. A connection between the DOT1L interacting partner ZFP-1 (AF10 in mammals) and RNAi in C. elegans was suggested by genetic experiments 13-15 years ago. To understand this connection, my laboratory undertook molecular and genomic studies of ZFP-1 and DOT1L. These studies not only confirmed the connection to RNAi, but also provided novel insights about the conserved role of DOT1L in transcription regulation. The DOT1L enzyme is essential for mammalian development and has been implicated in the pathogenesis of leukemia and breast cancer. However, the exact role of DOT1L in gene regulation is not clear. Preliminary work from our lab has begun to reveal this role; we have discovered that DOT1L and H3K79 methylation globally suppress non-coding RNA transcription in C. elegans, including that of antisense and enhancer RNAs. Our preliminary data strongly suggest ectopic dsRNA formation and small RNA production upon DOT1L loss-of-function. Our main goals are: 1) to detect and characterize these dsRNA and small RNA molecules in C. elegans; 2) to explore the possibility that the dsRNA suppressing function of DOT1L is conserved in mammals, and 3) to investigate the mechanism of transcription regulation by DOT1L in development using the C. elegans and zebrafish systems.

我们研究的广泛目标是了解动物发育过程中如何调节基因表达。我们的具体兴趣是找到内源性双链RNA产生的自然调节剂和核RNA干扰。在这方面，我们专注于RNA干扰与组蛋白H3赖氨酸79 DOT1L甲基转移酶复合物。 DOT1L交互之间的连接伴侣ZFP-1（哺乳动物中的AF10）和秀丽隐杆线虫中的RNAi通过遗传实验13 - 15年。前。为了了解这一联系，我的实验室进行了ZFP-1和 dot1l。这些研究不仅证实了与RNAi的联系，而且还提供了有关该研究的新见解 DOT1L在转录调控中的保守作用。 DOT1L酶对于哺乳动物至关重要发育并与白血病和乳腺癌的发病机理有关。但是，确切的 DOT1L在基因调节中的作用尚不清楚。我们实验室的初步工作已经开始揭示这一角色。我们已经发现DOT1L和H3K79甲基化全球抑制了C中的非编码RNA转录。秀丽隐杆线，包括反义和增强剂RNA。我们的初步数据强烈建议异位dsrna DOT1L功能丧失后的形成和小RNA产生。我们的主要目标是：1）检测和表征秀丽隐杆线虫中的这些dsRNA和小的RNA分子； 2）探索dsRNA的可能性 DOT1L的抑制功能在哺乳动物中是保守的，3）研究转录机理使用秀丽隐杆线虫和斑马鱼系统在开发中对DOT1L进行调节。