Epigenetic and post-translational modifier regulation of juvenile hormone action

保幼激素作用的表观遗传和翻译后修饰调节

• 批准号: 9028572
• 负责人: SUBBA R PALLI
• 金额: $ 28万
• 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 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9028572
• 关键词: Acetylation; Adult; Agonist; Biological; Biological Assay; Biological Metamorphosis; Biological Models; CHD1 gene; Cell Culture Techniques; Cells; Chromatin; Culicidae; Cyclic AMP-Responsive DNA-Binding Protein; DNA; Development; Disease; Disease Vectors; Ecdysone; Ecdysteroids; Ensure; Epigenetic Process; Event; Food; Gene Expression Regulation; Genes; Helix-Turn-Helix Motifs; Histone Acetylation; Histone Deacetylase; Histone Deacetylase Inhibitor; Histone H3; Histones; Homologous Gene; Hormone Receptor; Hormones; Human; Insect Control; Insect Vectors; Insecta; Insecticides; Juvenile Hormones; Knowledge; Laboratories; Lead; Livestock; Lysine; Mass Spectrum Analysis; Mediating; Medicine; Methods; Methoprene; Molecular; Molting; Nuclear Proteins; Outcome Study; Physiological; Play; Post-Translational Protein Processing; Post-Translational Regulation; Proteins; RNA Interference; Regulation; Reporter; Reproduction; Research; Role; Steroid Receptors; Testing; Tissues; Transferase; Tribolium; Trichostatin A; Vertebrates; base; brahma; genome sequencing; hormone regulation; human disease; method development; novel; promoter; public health relevance; receptor; research study; response; success; transcription factor; vector control; whole genome; yeast two hybrid system

 DESCRIPTION (provided by applicant): There has been a continuous demand for the development of insect control methods that are target-specific. Juvenile hormone (JH) and ecdysteroids are the major hormones that regulate development and reproduction in insects. Since these hormones are not present in vertebrates, they represent attractive targets for the development of insect control methods. Hindering this effort is the lack of understanding on the molecular basis of JH action. Tremendous progress has been made in understanding JH action at the molecular level during the past few years. A bHLH transcription factor, methoprene tolerant (Met), has been identified as a JH receptor. A steroid receptor co-activator homologue (SRC/Taiman/FISC) and Cycle have been identified as important co-factors involved in JH action. Kruppel homologue (Kr-h1) and bHLH transcription factor, Hairy and E93 have been identified as important transcription factors that mediate JH action. Preliminary experiments showed that Cyclic AMP response-element binding protein (CBP) is required for acetylation of histone H3 as well as for JH induction of JH-response genes in Tribolium castaneum tissues, TcA and Aag-2 cells. In addition, histone deacetylase (HDAC) inhibitor, Trichostatin A (TSA) also induces expression of JH-response genes in TcA and Aag-2 cells. Chromatin modifiers Brahma, Snr1 and CHD1 but not DNA methyl transferase are required for regulation of metamorphosis and reproduction in T. castaneum. JH exerts diverse functions in different tissues and under various physiological conditions. We hypothesize that epigenetic and post- translational modification of proteins involved in JH action play critical roles in multi-dimensionl gene regulation strategies employed by JH. Therefore, the main objective of this proposal is to capitalize on these latest developments to identify epigenetic modifiers and determine their mechanisms of action in modulation of JH response using T. castaneum, TcA and Aag-2 cells. The two Specific Aims of the proposal are 1. To identify key players involved in epigenetic and post-translational modifier regulation of JH action and 2. To determine the mechanisms involved in epigenetic and post-translational modifier regulation of JH action. In the first Specific Aim, w will employ RNAi, cell culture and reporter assays to identify and characterize key proteins including histone acetyl transferases (HAT), HDACs and chromatic modifiers involved in modulation of JH action. Temporal, spatial and hormonal regulation of identified genes as well as their function in JH action will be determined. In the second Specific Aim, we will employ mass spectrometry, RNAi, two-hybrid, pull- down, Chip and reporter assays to identify acetylated lysines in Met, domains of HATs, HDACs and Brahma, Snr1 and CHD1 required for JH action, localize HATs, HDACs and Brahma, Snr1 and CHD1 on Kr-h1, Hairy and E93 promoters and identify interacting partners of HATs, HDACs, Brahma, Snr1 and CHD1. The expected outcome of this study, an understanding of the molecular basis of JH action and identification of critical proteins involved in JH action will help in development of methods for control of insect vectors. The proposed studies will also advance our knowledge on epigenetic and post-translational modification of histones and other nuclear proteins especially on the functions and mechanisms of action of the modifiers.

 描述（由申请人提供）：对于开发针对特定目标的昆虫控制方法的需求一直存在。保幼激素（JH）和蜕皮激素是调节昆虫发育和繁殖的主要激素。由于这些激素不存在于脊椎动物中，因此它们代表了昆虫控制方法开发的有吸引力的目标。阻碍这一努力的是缺乏对 JH 作用分子基础的了解。过去几年，在分子水平上理解 JH 作用方面取得了巨大进展。 bHLH 转录因子甲氧二烯耐受 (Met) 已被鉴定为 JH 受体。类固醇受体辅激活剂同源物 (SRC/Taiman/FISC) 和 Cycle 已被确定为参与 JH 作用的重要辅因子。 Kruppel 同源物 (Kr-h1) 和 bHLH 转录因子、Hairy 和 E93 已被确定为介导 JH 作用的重要转录因子。初步实验表明，在赤拟谷盗组织、TcA 和 Aag-2 细胞中，组蛋白 H3 乙酰化以及 JH 反应基因的 JH 诱导需要环状 AMP 反应元件结合蛋白 (CBP)。此外，组蛋白脱乙酰酶 (HDAC) 抑制剂曲古抑菌素 A (TSA) 也会诱导 TcA 和 Aag-2 细胞中 JH 反应基因的表达。栗色砗磲变态和繁殖的调节需要染色质修饰剂 Brahma、Snr1 和 CHD1，但不需要 DNA 甲基转移酶。 JH在不同的组织和不同的生理条件下发挥不同的功能。我们假设参与 JH 作用的蛋白质的表观遗传和翻译后修饰在 JH 采用的多维基因调控策略中发挥着关键作用。因此，本提案的主要目标是利用这些最新进展来识别表观遗传修饰剂并确定其在使用赤霉、TcA 和 Aag-2 细胞调节 JH 反应中的作用机制。该提案的两个具体目标是 1. 确定参与 JH 作用的表观遗传和翻译后修饰调节的关键参与者；2. 确定参与 JH 作用的表观遗传和翻译后修饰调节的机制。在第一个具体目标中，我们将采用 RNAi、细胞培养和报告分析来识别和表征关键蛋白，包括组蛋白乙酰转移酶 (HAT)、HDAC 和参与调节 JH 作用的染色修饰剂。将确定已识别基因的时间、空间和激素调节及其在 JH 作用中的功能。在第二个具体目标中，我们将采用质谱、RNAi、双杂交、pull-down、芯片和报告基因检测来鉴定 JH 作用所需的 Met、HAT 结构域、HDAC 和 Brahma、Snr1 和 CHD1 中的乙酰化赖氨酸，定位 Kr-h1、Hairy 和 E93 启动子上的 HAT、HDAC 和 Brahma、Snr1 和 CHD1，并识别 HAT、HDAC、Brahma、Snr1 和 CHD1。这项研究的预期结果，即了解 JH 作用的分子基础和鉴定参与 JH 作用的关键蛋白质将有助于开发昆虫媒介控制方法。拟议的研究还将增进我们对组蛋白和其他核蛋白的表观遗传和翻译后修饰的了解，特别是修饰剂的功能和作用机制。