Chromatin mechanism of molecular motor gene regulation

描述（由申请人提供）：病理性应激诱导心肌中的转录基因重编程，导致肌病和心力衰竭。然而，很少有人知道病理应激如何触发染色质的重组，以控制肌病基因的表达。该研究项目的重点是不同的染色质调节因子如何协同作用，以建立抑制性染色质环境来控制α-肌球蛋白重链（<$-MHC）的表达，这是一种分子马达基因，其在应激心脏中的抑制导致肌病和心力衰竭。四种不同的染色质调节染色质重塑，组蛋白甲基转移酶，组蛋白脱乙酰酶和DNA甲基转移酶，将检查他们的合作抑制？-MHC。由于化学抑制剂可用于抑制这些染色质因子，因此有可能开发drus来增强衰竭心脏中的<$- MHC以改善心脏功能。了解这些染色质因子（药物靶标）如何相互作用将有助于我们设计新药，并了解抑制不同类型染色质调节剂的潜在药物的协同作用/动力学。目的1：确定特定亚型的DNA甲基转移酶所需的<$-MHC抑制和心力衰竭。我们将敲除小鼠心肌中的DNA甲基转移酶亚型，并检查其对应激心脏中的<$-MHC抑制和心力衰竭进展的影响。目的2：确定染色质重塑如何整合组蛋白和DNA甲基转移酶沉默？-MHC。我们将使用动物模型，手术方法和各种分子生物学方法（染色质免疫沉淀和定量PCR）来确定这些染色质因子之间的相互作用，这些染色质因子对组蛋白和<$-MHC的DNA甲基化至关重要。目的3：确定组蛋白去乙酰化酶如何与组蛋白和DNA甲基转移酶协同抑制<$-MHC启动子。我们将使用动物模型，手术方法和各种分子生物学方法（染色质免疫沉淀，定量PCR和免疫共沉淀）来定义这些染色质因子之间的相互作用，这些染色质因子对组蛋白甲基化和乙酰化以及<$-MHC启动子的DNA甲基化至关重要。

DESCRIPTION (provided by applicant): Pathological stress induces transcriptional gene reprogramming in the heart muscle, leading to myopathy and heart failure. However, little is known about how the pathological stress triggers the restructuring of chromatin to control myopathic gene expression. This research program focuses on how different chromatin-regulating factors act in concert to establish a repressive chromatin environment to control the expression of alpha-myosin heavy chain (¿-MHC), a molecular motor gene whose repression in the stressed hearts contributes to myopathy and heart failure. Four different chromatin regulators-chromatin remodeler, histone methyltransferase, histone deacetylase and DNA methyltransferase-will be examined for their collaborative repression of ¿-MHC. Because chemical inhibitors are available to inhibit these chromatin factors, it is possible to develop drus to boost ¿- MHC in the failing heart to improve cardiac function. Understanding how these chromatin factors (drug targets) interact will help us design new drugs and understand the synergy/dynamics of potential drugs that inhibit different classes of chromatin regulators. Aim 1: Defining specific isoforms of DNA methyltransferase required for ¿-MHC repression and heart failure. We will knock out DNA methyltransferase isoforms in the heart muscle of mice and examine the effects on ¿-MHC repression in the stressed heart and on the progression of heart failure. Aim 2: Determine how chromatin remodeler integrates histone and DNA methyltransferases to silence ¿-MHC. We will use animal models, surgical method, and various molecular biology methods (chromatin immunoprecipitation and quantitative PCR) to define the interactions between these chromatin factors crucial for the histone and DNA methylation of ¿-MHC. Aim 3: Defining how histone deacetylase coordinates with histone and DNA methyltransferases to repress ¿-MHC promoter. We will use animal models, surgical methods, and various molecular biology methods (chromatin immunoprecipitation, quantitative PCR, and co-immunoprecipitation) to define the interactions between these chromatin factors that are essential for histone methylation and acetylation, as well as DNA methylation of ¿-MHC promoter.