The role of H3K27me2 in regulating Culex pipiens diapause

项目摘要 昆虫经历各种进化过程才能在极端环境中生存 和气候。滞育是许多昆虫适应环境的最有效手段之一 温和的环境我们的目标是确定H3 K27 me 2是否调节 滞育特征包括寿命延长、脂肪积累和压力增强 北方家蚊Cx. pipiens。我们有三个具体目标， 本研究的主要内容如下：1）建立滞育相关基因与H3 K27 me 2的关联 脂肪体组织的损失。我们将确定H3 K27 me 2的遗传靶点， 利用三种功能测定评估基因在滞育性状中的假定作用：ChIP- seq、RNA-seq和RNA干扰（RNAi）。2)为了识别和描述这些基因 通过H3 K27 me 2损失激活以产生越冬脂肪储存， 消费我们将确定脂肪体细胞中的几十种特定类型的脂质 利用SSNMR和FTIR方法研究了越冬期间的土壤水分变化。第二，我们将使用 KEGG Mapper预测更高水平的生化和功能途径， 在暴露于滞育诱导条件的脂肪体组织中差异表达。 3)定义组蛋白去甲基化酶（UTX）和组蛋白 甲基转移酶（E（z）和ESC）在调节H3 K27 me 2中的作用，以了解 延长滞育蚊子寿命的机制。我们将描述 utx、e（z）和esc在调节H3 K27甲基化和滞育寿命中的作用 女性CX pipiens。这将通过敲下这些表达式来实现 基因或注射针对编码蛋白质的化学抑制剂。我们将 监测对H3 K27甲基化水平和患者存活率的影响。 处理Cx。pipiens。组蛋白甲基化可能是破坏或 逆转滞育相关的生理变化。像其他表观遗传 由于组蛋白甲基化是可逆的，因此通过改变组蛋白甲基化的活性， 酶调节这些修改，它可能会开发一种新的控制 策略，以延迟或防止这些蚊子越冬滞育的发病 从而在冬季造成大量的种群损失。

PROJECT SUMMARY Insects go through various evolutionary processes to survive in extreme environments and climates. Diapause is one of the most effective means that many insects adapted for a temperate environment. Our objective is to determine whether H3K27me2 regulates diapause traits including extended lifespan, fat accumulation, and enhanced stress resistance in the northern house mosquito, Cx. pipiens. We have three specific goals for this project: 1) To establish the link between diapause-relevant genes and H3K27me2 loss in fat body tissues. We will identify the genetic targets of the H3K27me2 and evaluate the genes’ putative roles in diapause traits using three functional assays: ChIP- seq, RNA-seq, and RNA interference (RNAi). 2) To identify and characterize the genes that are activated by H3K27me2 loss to generate the overwintering fat storage and consumption. We will identify dozens of specific types of lipids in the fat body cells during the overwintering periods using SSNMR and FTIR methods. Second, we will use KEGG Mapper to predict higher-level biochemical and functional pathways that are differentially expressed in fat body tissues exposed to the diapause-inducing condition. 3) To define the roles of the histone demethylase (UTX) and the histone methyltransferases (E(z) and ESC) in regulating H3K27me2 in order to understand the mechanisms that extend the lifespan of diapausing mosquitoes. We will characterize the role of utx, e(z) and esc in modulating H3K27 methylation and lifespan of diapausing females of Cx. pipiens. This will be achieved by knocking down the expression of these genes or by injecting chemical inhibitors that target the encoded proteins. We will monitor the impact on the H3K27 methylation levels and on the survival rates of the treated Cx. pipiens. Histone methylation is potentially a powerful target for breaking or reversing diapause-associated physiological changes. Like other epigenetic modifications, histone methylation is reversible, and thus by altering the activity of the enzyme that regulates these modifications, it may be possible to develop a novel control strategy to delay or prevent the onset of overwintering diapause in these mosquitoes and thus cause significant population loss during the winter season.