Homeostatic Roles of Connexin43 in Response to DNA Damage

Connexin43 在 DNA 损伤反应中的稳态作用

• 批准号: 8044043
• 负责人: Daniel Sam Roh
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044043
• 关键词: Affect; Aging; Aging-Related Process; Antibodies; Apoptosis; Biological Assay; Cell Communication; Cell Proliferation; Cell Survival; Cell physiology; Cells; Cessation of life; Connexin 43; Connexins; Cornea; Corneal Endothelium; DNA Damage; DNA Repair; Data; Detergents; Deterioration; Endothelial Cells; Epithelium; Event; Exposure to; Future; Gap Junctions; Genes; Genotoxic Stress; Goals; Homeostasis; In Vitro; Intercellular Communication Inhibition; Lead; Link; Maintenance; Mediating; Modification; Molecular; Mus; Organ; Permeability; Phosphorylation; Phosphotransferases; Physiological; Proteins; Pump; RNA Interference; Resistance; Role; Signal Transduction; Site; Stress; Testing; Tissues; age related; anterior chamber; biological adaptation to stress; casein kinase; casein kinase I; cell injury; coping; coping mechanism; driving force; in vitro Model; in vivo; inhibitor/antagonist; insight; intercellular communication; lens; neuronal cell body; response; stressor; trafficking

DESCRIPTION (provided by applicant): DNA damage can significantly alter cell function and viability, thus, the accumulation of unrepaired DNA damage is believed to be a major influence in the deterioration of organ function in aging. However, cells also respond to DNA damage in many other ways beyond DNA repair in order to maintain their function and viability. In particular, genes involved in intercellular communication, such as connexins, have been identified as key components in cellular responses to numerous stressors. Despite this, the molecular mechanisms by which intercellular communication is modified after genotoxic stress have not been described. The overall goal of this F30 proposal is to understand the responses to DNA damage in corneal endothelial (CE) cells whose pump and barrier functions are essential for corneal transparency and which in vivo display age-related degeneration and accumulation of DNA damage. Using an in vitro model of CE cells, we have recently observed significant changes in the gap junction protein connexin-43 (Cx43) after exposure to DNA damage-inducing agents. To explain the cellular mechanism(s) behind these changes, we consider a potential link between DNA damage and cell communication via casein kinase-1 delta (CK1?), which is both activated by DNA damage and an essential Cx43 kinase. Our overall hypothesis is that a protective cellular response to DNA damage involves stabilization of gap junction intercellular communication which is (a) mediated by changes in Cx43 expression and site-specific phosphorylation by CK1?), and (b) required for maintenance of viability and function. This will be tested with two specific aims that will determine 1) the mechanism(s) by which DNA damage mediates modification of connexin-43 (Cx43) and 2) the physiological consequence(s) of these changes during DNA damage including effects of CE cell viability and function. The results of this project will further our understanding of stress responses to DNA damage and provide insights into how the viability of certain aging tissues may be potentially enhanced. The aging process in which the body's cells, tissues, and organs progressively deteriorate has been associated with the accumulation of DNA damage. In response to such stress, a cell may survive or perish depending on its ability to cope with that damage. This proposal studies a potential coping mechanism to DNA damage that involves changes in how damaged cells communicate with one another to maintain their function and viability.

描述（由申请人提供）：DNA损伤可以显着改变细胞功能和生存能力，因此，未修复的DNA损伤的积累被认为是衰老中器官功能恶化的主要影响。但是，细胞还以除DNA修复以外的许多其他方式对DNA损伤做出反应，以保持其功能和生存能力。特别是，涉及细胞间通信的基因，例如连接素，已被鉴定为对众多胁迫响应的细胞反应中的关键组成部分。尽管如此，尚未描述遗传毒性应激后修饰细胞间通信的分子机制。该F30提案的总体目标是了解角膜内皮（CE）细胞中对DNA损伤的反应，其泵和屏障功能对于角膜透明度至关重要，并且在体内显示与年龄相关的变性和DNA损伤的积累。使用CE细胞的体外模型，我们最近观察到暴露于DNA损伤剂后的间隙连接蛋白连接素-43（CX43）的显着变化。为了解释这些变化背后的细胞机制，我们考虑了通过酪蛋白激酶-1 Delta（CK1？）之间的DNA损伤与细胞通信之间的潜在联系，这两者都被DNA损伤激活，而必需的CX43激酶。我们的总体假设是，对DNA损伤的保护性细胞反应涉及间隙连接间的细胞间通信的稳定，（a）是由CX43表达的变化和CK1的位点特异性磷酸化介导的，并且（b）维持生存能力和功能所需。这将通过两个具体目标对1）DNA损伤介导连接蛋白43（CX43）和2）在DNA损伤过程中这些变化的生理后果的机制进行测试，以确定1）DNA损伤的机制。该项目的结果将进一步了解对DNA损伤的压力反应，并提供有关如何有可能增强某些衰老组织的生存能力的见解。人体细胞，组织和器官逐渐恶化的衰老过程与DNA损伤的积累有关。为了应对这种压力，细胞可能会根据应对该损害的能力而生存或灭亡。该建议研究了一种潜在的应对机制，以涉及涉及损坏细胞彼此沟通以保持其功能和生存能力的变化。