权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Gene Regulation by and of LEDGF

LEDGF 的基因调控

基本信息

批准号：
8204533
负责人：
DHIRENDRA P SINGH
金额：
$ 35.28万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-03-01 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8204533
关键词：
Acquired Immunodeficiency Syndrome Affect Affinity Age Aging Amino Acids Antibodies Apoptosis Attenuated Autoimmune Process Binding Bioinformatics Biological Assay C-terminal Cataract Cell Death Cell Survival Cell physiology Cells Cellular Stress Chronic stress Collaborations Competence Complementary DNA Coupled Crystallins Cytoprotection DNA Binding DNA Binding Domain DNA-Protein Interaction Data Deletion Mutation Disease Drug Formulations Electrophoretic Mobility Shift Assay Elements Epithelial Cells Figs - dietary Funding Future Gene Expression Gene Expression Regulation Genes Genetic Transcription Goals Grant Growth Growth Factor Growth Factor Gene Heat Stress Disorders Heat shock proteins Heat-Shock Proteins 70 Heat-Shock Response Heat-Shock Transcription Factor 2 Heating Helix-Turn-Helix Motifs Homeostasis Impairment Investigation Letters Literature Malignant Neoplasms Maps Measures Mediating Methods Modification Molecular Molecular Structure Monitor Mutagenesis Mutate N-terminal Nature Oxidative Stress Peptide Hydrolases Phosphorylation Phosphorylation Site Physiological Physiological Processes Point Mutation Post-Translational Protein Processing Predisposition Principal Investigator Process Proteins Published Comment Publishing Qualifying RNA Reaction Reagent Reporting Research Research Personnel Resistance Resources Role Running Sampling Signal Transduction Site Site-Directed Mutagenesis Small Interfering RNA Stress Stretching Structure-Activity Relationship Suggestion System TNF gene Therapeutic Time Transactivation Transfection Ubiquitin Work acute stress age related cell type combat experience expression vector gel mobility shift assay gene interaction heat-shock factor 1 insight interest knock-down lens lens epithelium-derived growth factor mutant novel overexpression programs promoter protein function protein protein interaction research study response satisfaction stoichiometry stress protein transcription factor vector yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Environmental stress is a major cause of age related cataract (ARC). Acute or chronic stresses provoke multiple cellular reactions, including activation of the heat shock or stress protein response. Lens epithelial cell-derived growth factor (LEDGF) is a stress-inducible transcription factor having pro-survival, cytoprotective function. Overall, LEDGF achieves this function by transactivating the stress-associated genes such as heat shock protein genes, for which LEDGF binds to the heat shock element (HSE) and stress element (STRE) present in the promoter of these genes. The specific molecular mechanism(s) by which LEDGF exerts its cytoprotective action under cellular stress is poorly understood, but various possibilities may exist, such as protein-protein and protein-DNA interactions involving LEDGF, its post-translational modifications, and the level of LEDGF gene expression. Our preliminary data show that (a) LEDGF binds with trimeric HSF1 during stress that cooperatively transactivates the heat shock genes - an interaction that leads cytoprotection; (b) LEDGF and HSF1 interaction is essential for cell survival under stress, as Hsf1 depleted cells, despite displaying higher LEDGF expression, are more prone to stress-induced cell death than are wild type cells; and (c) SUMO-1 modification of LEDGF, a transcription factor, and sumoylation of transcriptional proteins leads to gene modulation; a mechanism involved in cellular signaling. These data suggest the existence of molecular interaction between LEDGF and HSF1. More specifically, how and when these two transcriptional proteins interact, how LEDGF expression is upregulated, and how its sumoylation affects transcription of heat shock genes remain to be understood. Therefore, we hypothesize that during stress and/or normal physiological conditions, LEDGF, a multidomain protein, exerts its cytoprotective role through multiple modes of action, viz. by DNA binding activity, protein modification by sumoylation and protein-protein interaction, and by its expression levels. The following specific aims will be pursued: 1) to map and determine the sites of SUMO-1 modification in LEDGF protein and to define its functional consequences for the DNA-binding and transactivation activity of LEDGF, 2) to define and characterize interacting domains of LEDGF and HSF1 and determine underlying mechanism(s) of their interaction during stress, 3) to demonstrate the effect of LEDGF and HSF1 interaction and/or their sumoylation in transactivation of heat shock genes, hsp27 or 1B-crystallin, and 4) to demonstrate that LEDGF and HSF1 interaction is necessary for stress-induced cellular cytoprotection. These studies will reveal new information concerning the molecular modes of LEDGF function under cellular stress in lens epithelial cells leading to future formulation of therapeutic strategy to combat age- related impairment of lens function, and other disorders in general such as Cancer and AIDS. Cloned LEDGF is a novel growth and survival factor, and acts as transcription factor. Despite the known regulatory function of LEDGF, only limited investigation has been made into the molecular structure-function relationship of LEDGF's domains. The research proposed in current grant will disclose the functionality of LEDGF domains; that will provide important insights into how LEDGF functions at the cellular and molecular levels, how it regulates cellular survival and other physiological processes, how these processes go awry in various disease states, and how the expression levels of LEDGF are regulated in cells facing stress. Thus accomplishing these goals in turn will provide opportunity to manipulate this molecule to cure and control various age associated diseases including cataract as well as other diseases, such as cancer, AIDS and autoimmune disoders, in general.

描述（由申请人提供）：环境压力是年龄相关性白内障（ARC）的主要原因。急性或慢性应激会引发多种细胞反应，包括激活热休克或应激蛋白反应。晶状体上皮细胞衍生生长因子（LEDGF）是一种应激诱导转录因子，具有促生存、细胞保护功能。总体而言，LEDGF通过反式激活应激相关基因（例如热休克蛋白基因）来实现这一功能，其中LEDGF与这些基因启动子中存在的热休克元件（HSE）和应激元件（STRE）结合。 LEDGF 在细胞应激下发挥细胞保护作用的具体分子机制尚不清楚，但可能存在各种可能性，例如涉及 LEDGF 的蛋白质-蛋白质和蛋白质-DNA 相互作用、其翻译后修饰以及 LEDGF 基因表达水平。我们的初步数据表明，(a) LEDGF 在应激过程中与三聚体 HSF1 结合，协同反式激活热休克基因——这种相互作用导致细胞保护； (b) LEDGF和HSF1相互作用对于细胞在应激下生存至关重要，因为Hsf1耗尽的细胞尽管表现出较高的LEDGF表达，但比野生型细胞更容易发生应激诱导的细胞死亡； (c) LEDGF（一种转录因子）的 SUMO-1 修饰以及转录蛋白的 sumoy 化导致基因调节；参与细胞信号传导的机制。这些数据表明 LEDGF 和 HSF1 之间存在分子相互作用。更具体地说，这两种转录蛋白如何以及何时相互作用、LEDGF 表达如何上调以及其苏酰化如何影响热休克基因的转录仍有待了解。因此，我们假设在应激和/或正常生理条件下，LEDGF（一种多结构域蛋白）通过多种作用模式发挥其细胞保护作用，即：通过 DNA 结合活性、通过苏酰化和蛋白质-蛋白质相互作用进行的蛋白质修饰及其表达水平。将追求以下具体目标：1) 绘制和确定 LEDGF 蛋白中 SUMO-1 修饰的位点，并定义其对 LEDGF 的 DNA 结合和反式激活活性的功能影响，2) 定义和表征 LEDGF 和 HSF1 的相互作用域，并确定它们在应激期间相互作用的潜在机制，3) 证明 LEDGF 和 HSF1 相互作用和/或它们的作用的影响。热休克基因、hsp27 或 1B-晶状体蛋白反式激活中的 sumoylation，以及 4) 证明 LEDGF 和 HSF1 相互作用对于应激诱导的细胞保护是必需的。这些研究将揭示有关晶状体上皮细胞细胞应激下 LEDGF 功能的分子模式的新信息，从而导致未来制定治疗策略，以对抗与年龄相关的晶状体功能损伤以及癌症和艾滋病等其他疾病。克隆的 LEDGF 是一种新型生长和存活因子，并充当转录因子。尽管 LEDGF 的调节功能已知，但对 LEDGF 结构域的分子结构与功能关系的研究却很有限。当前资助中提出的研究将揭示 LEDGF 域的功能；这将为了解 LEDGF 如何在细胞和分子水平上发挥作用、如何调节细胞生存和其他生理过程、这些过程在各种疾病状态下如何出错以及面临压力的细胞中 LEDGF 的表达水平如何受到调节提供重要见解。因此，实现这些目标反过来将提供操纵该分子来治疗和控制各种与年龄相关的疾病的机会，包括白内障以及其他疾病，例如癌症、艾滋病和自身免疫性疾病。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Peroxiredoxin 6 delivery attenuates TNF-alpha-and glutamate-induced retinal ganglion cell death by limiting ROS levels and maintaining Ca2+ homeostasis.

DOI：
10.1016/j.brainres.2008.07.076
发表时间：
2008-10-03
期刊：
Brain research
影响因子：
2.9
作者：
Fatma N;Kubo E;Sen M;Agarwal N;Thoreson WB;Camras CB;Singh DP
通讯作者：
Singh DP

Dynamic and differential regulation in the microRNA expression in the developing and mature cataractous rat lens.

DOI：
10.1111/jcmm.12094
发表时间：
2013-09
期刊：
Journal of cellular and molecular medicine
影响因子：
5.3
作者：
Kubo E;Hasanova N;Sasaki H;Singh DP
通讯作者：
Singh DP

Lens epithelium-derived growth factor deSumoylation by Sumo-specific protease-1 regulates its transcriptional activation of small heat shock protein and the cellular response.

DOI：
10.1111/j.1742-4658.2012.08686.x
发表时间：
2012-09
期刊：
The FEBS journal
影响因子：
0
作者：
Ishihara K;Fatma N;Bhargavan B;Chhunchha B;Kubo E;Dey S;Takamura Y;Kumar A;Singh DP
通讯作者：
Singh DP

Neuroprotective effect of peroxiredoxin 6 against hypoxia-induced retinal ganglion cell damage.