Metabolic control of global gene expression during the Maternal-to-Zygotic Transition

母体向合子转变期间整体基因表达的代谢控制

• 批准号: 10542564
• 负责人: Lital Bentovim
• 金额: $ 3.82万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10542564
• 关键词: ATAC-seq; Animal Model; ChIP-seq; Chromatin; DNA; Data; Development; Developmental Gene; Developmental Process; Diabetes Mellitus; Disease; Drosophila genus; Embryo; Enzymes; Gene Abnormality; Gene Activation; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomic approach; Global Change; Goals; Histones; Human; Image; Lead; Link; Malignant Neoplasms; Mass Spectrum Analysis; Maternal Messenger RNA; Measures; Messenger RNA; Metabolic; Metabolic Control; Metabolic dysfunction; Metabolism; Methods; Methylation; Modification; Molecular Genetics; Paint; Play; Process; RNA; RNA Interference; Regulation; Reporter; Research; Research Personnel; Resolution; Role; Shapes; Signal Transduction; System; Testing; Therapeutic Intervention; Time; Transcript; Transcriptional Activation; Western Blotting; Zebrafish; alpha ketoglutarate; demethylation; egg; experimental study; genetic approach; histone methylation; histone modification; human disease; in vivo; mRNA Decay; mRNA Transcript Degradation; metabolic abnormality assessment; mutant; novel; programs; skills; targeted treatment; tool; transcriptome; transcriptome sequencing

ABSTRACT After an egg is fertilized, many thousands of genes are turned on in the embryo while maternal mRNA is degraded; this process is called the maternal-to-zygotic transition (MZT). The goal of this proposal is to decipher how activation of zygotic genes is synchronized with maternal mRNA decay. Both histone modifications and the mRNA modification N6-methyladenosine (m6A) play prominent roles, during the MZT, by regulating transcription of zygotic genes and maternal mRNA clearance, respectively. However, the signals that control changes in chromatin and mRNA modifications in this critical developmental stage are largely unknown. This proposal will test the novel hypothesis that changes in cellular metabolism coordinate global gene expression during the MZT, through modulating the availability of metabolites required for chromatin and mRNA modifications. Using high-resolution mass-spectrometry, in Drosophila (D). melanogaster embryos during the MZT, we identified changes in the levels of alpha-ketoglutarate (α-KG), which is a required co-factor for histone and RNA demethylation enzymes. Here, we test our central hypothesis through the following specific aims: (1) test the link between α-KG levels, histone methylation, and global transcription activation of zygotic genes, and (2) test the link between α-KG levels, m6A modification of mRNA, and maternal mRNA decay. D. melanogaster embryos are an ideal system for studying the interplay between metabolism and gene regulation because there are many experimental tools for manipulating and measuring metabolism, chromatin state, mRNA modifications and transcription in vivo. This proposal will apply molecular, genetic and genomic approaches to demonstrate how metabolic inputs, such as α-KG, shape the developmental transcriptome and coordinate critical developmental processes by controlling both chromatin and mRNA modifications. Findings from this study will be relevant for understanding not only developmental processes but also diseases in which metabolic dysfunction is associated with abnormal gene expression, such as cancer and diabetes. Establishing D. melanogaster as an experimental platform for studying metabolic regulation of global gene expression and acquiring the relevant set of skills as part of this research plan will be a critical next step towards fulfilling my goal of becoming an independent investigator.

摘要 卵子受精后，胚胎中成千上万的基因被打开，而母体mRNA则被激活。 退化;这个过程被称为母亲到合子过渡（MZT）。本提案的目的是 解释合子基因的激活是如何与母体mRNA的衰变同步的。组蛋白 修饰和mRNA修饰N6-甲基腺苷（m6 A）在MZT期间起着重要作用， 通过调节合子基因的转录和母体mRNA的清除。然而，信号 在这个关键的发育阶段，控制染色质变化和mRNA修饰的基因主要是 未知该提案将检验细胞代谢变化协调全球的新假设 MZT期间的基因表达，通过调节染色质所需的代谢产物的可用性， 和mRNA修饰。使用高分辨率质谱，在果蝇（D）。melanogaster 在MZT期间，我们确定了α-酮戊二酸（α-KG）水平的变化，这是一个必需的 组蛋白和RNA去甲基化酶的辅助因子。在这里，我们通过 以下具体目标：（1）测试α-KG水平，组蛋白甲基化和整体转录之间的联系 合子基因的激活，和（2）测试α-KG水平，mRNA的m6 A修饰， 母体mRNA衰变。D.黑腹胚胎是研究 代谢和基因调控，因为有许多实验工具来操纵和测量 代谢、染色质状态、mRNA修饰和体内转录。这项建议将适用于 分子，遗传和基因组方法来证明代谢输入，如α-KG，如何塑造 发育转录组和协调关键的发育过程，通过控制染色质 和mRNA修饰。这项研究的结果不仅有助于了解发展 过程，而且还有代谢功能障碍与异常基因表达相关的疾病，如 癌症和糖尿病一样。建立D.作为研究代谢的实验平台 全球基因表达的调控和获得相关的一套技能，作为这项研究计划的一部分， 这是实现我成为独立调查员目标的关键一步。