The role of DOT1L methyltransferase in controlling the noncoding transcriptome

我们研究的广泛目标是了解基因表达在动物发育过程中是如何调节的。 我们特别感兴趣的是寻找内源性双链RNA生产和核的天然调节器 RNA干扰。在这方面，我们正在重点研究RNA干扰和组蛋白之间的关系 H3赖氨酸79 DOT1L甲基转移酶复合体。DOT1L交互伙伴ZFP-1之间的连接 (哺乳动物中的AF10)和线虫中的RNAi是13-15年前的遗传实验提出的。至 为了了解这种联系，我的实验室进行了ZFP-1和DOT1L的分子和基因组研究。 这些研究不仅证实了与RNAi的联系，而且也为保守的 DOT1L在转录调控中的作用。DOT1L酶对哺乳动物的发育是必不可少的，并具有 与白血病和乳腺癌的发病机制有关。然而，DOT1L在基因中的确切作用 监管尚不明确。我们实验室的初步工作已经开始揭示这种作用；我们已经发现 DOT1L和H3K79甲基化全局抑制线虫非编码RNA转录，包括线虫 反义和增强子RNA。我们的初步数据强烈表明异位dsrna的形成和小rna。 在DOT1L功能丧失时进行生产。我们的主要目标是：1)检测和鉴定这些dsRNA和 线虫中的小RNA分子；2)探讨DOT1L的dsRNA抑制功能的可能性 在哺乳动物中保守，以及3)研究DOT1L在哺乳动物中转录调节的机制 利用线虫和斑马鱼系统进行开发。 该项目不是专门针对生物伦理研究或生物伦理能力建设的。本科和高中 学生将积极参与拟议的研究。

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. The project is not specific to bioethics research or capacity building in bioethics. Undergraduate and high school students will actively participate in the proposed research.