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（由适用提供）：对于特定于目标的绝缘控制方法的开发一直有不断的需求。青少年激素（JH）和雌激素是调节绝缘发育和繁殖的主要骑兵。由于这些骑兵不存在于脊椎动物中，因此它们代表了开发绝缘控制方法的有吸引力的靶标。阻碍了这一努力是缺乏对JH作用的分子基础的理解。在过去几年中，在理解分子水平的JH作用方面取得了巨大进步。 BHLH转录因子Method -od -parerant（Met）已被鉴定为JH受体。类固醇受体共激活器同源物（SRC/Taiman/fisc）和循环已被确定为参与JH作用的重要共同因素。 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，循环AMP响应元素结合蛋白（CBP）是组蛋白H3的乙酰化以及JH诱导的JH反应基因在TCA组织，TCA和AAG-2细胞中所必需的。此外，组蛋白脱乙酰基酶（HDAC）抑制剂Trichostatin A（TSA）还影响TCA和AAG-2细胞中JH-反应基因的表达。染色质修饰剂梵天，SNR1和CHD1而不是DNA甲基转移是需要调节T. castaneum中变形和繁殖的。 JH在不同的组织和各种物理条件下发挥潜水功能。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采用的多二键基因调控策略中起关键作用。因此，该提案的主要目的是利用这些最新的发展，以识别表观遗传修饰剂，并使用T. castaneum，TCA和AAG-2细胞来确定其在调节JH响应时的作用机理。该提案的两个具体目的是1。确定JH动作的表观遗传和翻译后修饰剂调节的关键参与者和2。确定JH动作的表观遗传和翻译后修饰剂调节所涉及的机制。在第一个特定目的中，W将使用RNAi，细胞培养和报告基因测定法来识别和表征关键蛋白，包括组蛋白乙酰基转移（HAT），HDACS，HDAC和涉及调节JH作用调节的色度修饰符。将确定鉴定基因及其在JH作用中的功能的时间，空间和骑马调节。在第二个具体目标中，我们将采用质谱，RNAi，两杂交，下拉，芯片和记者测定法，以识别Met，Hats，HDACS和Brahma和Brahma的域，SNR1和CHD1，JH动作所需的乙酰化歌词帽子，HDAC，梵天，SNR1和CHD1。这项研究的预期结果，对JH作用的分子基础的理解以及对JH作用涉及的关键蛋白的鉴定将有助于开发用于控制绝缘载体的方法。拟议的研究还将提高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，尤其是在修饰剂作用的功能和机制方面。