Aldosterone-sensitive repression of ENaC by AF9 and Dot1

AF9 和 Dot1 对 ENaC 的醛固酮敏感抑制

• 批准号: 6909305
• 负责人: WENZHENG ZHANG
• 金额: $ 10.98万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6909305
• 关键词: acid base balance; aldosterone; binding sites; biological signal transduction; blood pressure; chromatin immunoprecipitation; electrolyte balance; gene induction /repression; genetic transcription; hormone regulation /control mechanism; kidney function; laboratory mouse; methylation; methyltransferase; molecular genetics; podocyte; polymerase chain reaction; protein protein interaction; protein structure function; sodium channel; transcription factor

DESCRIPTION (provided by applicant): The major objective of this research program is to provide the applicant with new and intensive training in relavant renal and whole animal physiological studies and a foundation of published work in elucidating aldosterone signaling pathways controlling ion transport criticle to renal physiology. The applicant's long-term goal is to become a successful independent investigator in molecular and cell biological research of the kidney. To achieve this goal, the applicant will be trained in the rich scientific environment of the Texas Medical Center under the close sponsorship of Dr. Bruce Kone, an expert in these areas. The scientific aims of the proposal are to model the molecular action of aldosterone on the epithelial Na+ channel in the collecting duct. Aldosterone is a major regulator of Na+ and acid-base balance and control of blood pressure. The specific aims are to define a novel signaling cascade initiated by aldosterone, via AF9a phosphorylation by SGK1 and H3 K79 methylation by mDotla to transcriptional activation of ENaC in mIMCDS cells and to reveal AF9a and mDotl a as novel physiological targets of aldosterone in regulating ENaC transcription in vivo in mice. mIMCDS cells have been shown to respond to aldosterone and express SGK1, AF9a, mDotl a and ENaC. A wide range of molecular approaches including chromatin immunoprecipitation, real-time RTqPCFt nuclear run-on assays and renal physiological assays (ion transport and urinary electrolyte) will be applied to achieve the goals. It is anticipated that 1) AF9a will identified as an early repressed target by aldosterone action, a novel physiological substrate of SGK1, the first non-histone protein interacting with mDotla and the first protein modulating H3 K79 methylation in a targeted manner; 2) Dotla will become the first chromatin-remodeling protein that serves as an integrate component of aldosterone signaling pathway; 3) ENaC will become the first immediate downstream target of AF9a. Collectively this work will poise the applicant to be an innovative independent scientist in renal signaling and ion transport.

描述（由申请人提供）： 该研究计划的主要目的是为申请人提供有关肾脏和整个动物生理学研究的新的，重密集型培训，并为阐明醛固酮信号传导途径的发表工作的基础，控制了离子运输批评至肾脏生理学。申请人的长期目标是成为肾脏分子和细胞生物学研究的成功独立研究者。为了实现这一目标，将在这些领域的专家布鲁斯·科恩（Bruce Kone）博士的密切赞助下，在得克萨斯医疗中心的丰富科学环境中接受培训。该提案的科学目的是对醛固酮在收集管中的上皮Na+通道上的分子作用进行建模。醛固酮是Na+和酸碱平衡和控制血压的主要调节剂。具体目的是定义醛固酮通过sgk1和H3 K79甲基化的AF9A磷酸化来定义一个新型的信号级联。已显示MIMCDS细胞对醛固酮和表达SGK1，AF9A，MDOTL A和ENAC反应。将应用各种分子方法，包括染色质免疫沉淀，实时RTQPCFT核跑步测定和肾脏生理测定（离子运输和尿液电解质）来实现目标。可以预料，1）AF9A将被醛固酮作用鉴定为早期抑制靶标，醛固酮作用是SGK1的新型生理底物，SGK1的新生理底物，第一种与MDOTLA相互作用的非轴承蛋白和第一种以靶向方式调节H3 K79甲基化的蛋白； 2）Dotla将成为第一个染色质复制蛋白，它是醛固酮信号通路的整合成分； 3）ENAC将成为AF9A的第一个立即下游目标。这项工作总的来说，将使申请人成为肾脏信号传导和离子运输方面的创新独立科学家。