Epigenetic regulation of hormone action in Tribolium and Aedes

谷盗和伊蚊激素作用的表观遗传调控

• 批准号: 10598572
• 负责人: SUBBA R PALLI
• 金额: $ 29.87万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10598572
• 关键词: Acetylation; Adult; Aedes; Agonist; Animals; Antibodies; Binding; Bioinformatics; Biological Assay; Biological Metamorphosis; Biological Models; CRISPR/Cas technology; Cell Culture Techniques; Cells; Complex; Culicidae; Data; Deacetylation; Development; Developmental Process; Disease; Disease Vectors; Ecdysone; Ecdysteroids; Ecdysterone; Embryonic Development; Endocrinology; Ensure; Environment; Enzymes; Epigenetic Process; Female; Food; Friends; Genes; Goals; Growth; Histone Deacetylase; Histone-Lysine N-Methyltransferase; Histones; Homologous Gene; Hormone Receptor; Hormones; Human; Insect Control; Insect Hormones; Insecta; Insecticides; Juvenile Hormones; Knowledge; Laboratories; Life; Ligands; Literature; Livestock; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Medicine; Methods; Methoprene; Methylation; Modeling; Modification; Molecular; Pathway interactions; Pest Control; Physiological; Physiological Processes; Plants; Play; Postembryonic; Principal Investigator; Process; Proteins; Publishing; RNA Interference; Regulation; Reporter; Reproduction; Reproductive Process; Research; Research Support; Response Elements; Role; Steroid Receptors; Testing; Tissues; Tribolium; Vertebrates; Western Blotting; chromatin immunoprecipitation; commercialization; demethylation; ecdysone receptor; enzyme activity; epigenetic regulation; experimental study; gene product; genome annotation; genome editing; histone acetyltransferase; histone demethylase; histone modification; hormone analog; hormone regulation; hormone response element; human disease; insect disease vector; mass spectrometer; programs; promoter; receptor; receptor binding; response; success; transcription factor; transcriptome sequencing; transmission process; vector control; yeast two hybrid system

PROJECT SUMMARY There has been a continuous demand for development of insect control methods that are target-specific. Juvenile hormone (JH) and ecdysteroids (20-hydroxyecdysone, 20E is the most active form) are the major hormones that regulate many physiological and developmental processes in insects. Since these hormones are not present in vertebrates, they represent attractive targets for development of insect control methods. Hindering this effort is the lack of understanding on the molecular basis of action of these hormones. There has been excellent recent progress in understanding the molecular basis of 20E and JH action. Key players, including receptors, co-activators, response elements and target genes involved in the action of these hormones have been identified. Non-steroidal ecdysone agonists and JH analogs have been discovered, and commercialized for the control of pests and disease vectors. Research supported by this program showed that epigenetic modifications, especially acetylation levels of histones regulated by histone acetyltransferases (HAT) and histone deacetylases (HDAC) modulate JH and 20E action. Hormone action requires multiple layers of regulation because the same circulating levels of hormones exert diverse functions in different tissues and under different physiological conditions. Based on the published literature and preliminary data included in this application, we hypothesize that epigenetic regulators HDACs, HATs, histone lysine methyltransferases (HKMT) and histone demethylases (HDM) modulate JH and 20E action in the regulation of postembryonic development and reproduction. The main goal of this proposal to test this hypothesis by studying the mechanisms of action of epigenetic modifiers in JH and 20E action using T. castaneum, TcA cells, Ae. aegypti and Aag2 cells as model systems. The two Specific Aims of the proposal are 1. To identify and characterize epigenetic modifiers required for 20E and JH regulation of postembryonic development and female reproduction and 2. To study mechanisms of action of epigenetic modifiers required for 20E and JH regulation of postembryonic development and female reproduction. In the first Specific Aim, RNAi, CRISPR/Cas9 genome editing methods, RNA sequencing, bioinformatics, RT-qPCR, enzyme assays and cell culture approaches will be used to identify and characterize HATs, HDACs, HKMTs and HDMs required for postembryonic development and female reproduction in T. castaneum and Ae. aegypti. In the second Specific Aim, the mechanisms of action of 10-12 epigenetic modifiers will be studied using western blots and ChIP, reporter, Two-hybrid and pull-down assays, and mass spectrometry approaches. Results from the proposed research will impact medicine by providing information for the development of vector control methods. This information could help in understanding epigenetics of human diseases such as cancer. The results from the proposed research may help to transform our understanding of endocrinology of disease vectors.

项目摘要 人们一直在不断开发目标特异性的昆虫控制方法的需求。 少年激素（JH）和ecdysteroid（20-羟基甲虫，20E是最活跃的形式）是主要的 调节昆虫中许多生理和发育过程的激素。因为这些激素 它们不存在于脊椎动物中，它们代表着开发昆虫控制方法的有吸引力的目标。 阻碍了这一努力是对这些激素作用的分子基础缺乏理解。有 在理解20E和JH作用的分子基础方面取得了绝佳的进步。主要参与者， 包括受体，共激活因子，响应元件和涉及这些作用的靶基因 已经确定了激素。已经发现了非甾体肌动力激动剂和JH类似物，并且 商业化以控制害虫和疾病媒介。该计划支持的研究表明 表观遗传修饰，尤其是组蛋白乙酰转移酶调节的组蛋白的乙酰化水平 （HAT）和组蛋白脱乙酰基酶（HDAC）调节JH和20E作用。激素作用需要多层 调节是因为相同的循环水平的激素在不同组织和 在不同的生理条件下。基于其中包括的文献和初步数据 应用，我们假设表观遗传调节剂HDAC，帽子，组蛋白赖氨酸甲基转移酶 （HKMT）和组蛋白脱甲基酶（HDM）调节JH和20E作用，以调节后胚后调节 开发和繁殖。该提议通过研究该假设的主要目标 使用T. castaneum，TCA细胞，AE，表观遗传修饰剂在JH和20E作用中的作用机理。埃及 和AAG2单元为模型系统。该提案的两个具体目的是1。 胚胎后发育和女性需要20E和JH调节所需的表观遗传修饰符 繁殖和2。研究20E和JH调节所需的表观遗传修饰剂的作用机理 胚胎后发育和女性繁殖。在第一个特定目标中，RNAi，crispr/cas9 基因组编辑方法，RNA测序，生物信息学，RT-QPCR，酶测定和细胞培养 方法将用于识别和表征帽子，HDAC，HKMT和HDMS所需的帽子， t. castaneum and Ae中的胚胎后发育和女性繁殖。埃及。在第二个特定 目的，将使用蛋白质印迹和芯片研究10-12个表观遗传修饰剂的作用机理， 记者，两杂交和下拉测定法以及质谱法。提议的结果 研究将通过提供向向量控制方法的开发信息来影响医学。这 信息可以帮助理解人类疾病（例如癌症）的表观遗传学。来自 拟议的研究可能有助于改变我们对疾病媒介内分泌学的理解。