Ethanol and MAP Kinase Cascade

乙醇和 MAP 激酶级联

• 批准号: 6733950
• 负责人: Shivendra D Shukla
• 金额: $ 26.05万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6733950
• 关键词: JUN kinase; SDS polyacrylamide gel electrophoresis; acute phase protein; alpha 1 acid glycoprotein; biological signal transduction; enzyme activity; enzyme induction /repression; ethanol; intracellular transport; isozymes; laboratory rat; liver cells; liver pharmacology; mitogen activated protein kinase; phospholipase A2; phosphoproteins; phosphorylation; polymerase chain reaction; protein metabolism; protein tyrosine kinase; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Molecular actions of ethanol remain largely unknown. Recent developments indicate that ethanol alters intracellular signaling pathways including those involving members of mitogen activated protein (MAP) kinase family. Based on new supporting data, it is hypothesized that "ethanol modulates p38 MAPK activity leading to downstream activation of nuclear mitogen and stress activated kinase-1 (MSK-1) and phosphorylation of histone-3 as target." An important focus of this renewal application is the ethanol effects on signaling in nucleus and its consequences, e.g., on histone modifications and on apoptosis. In these studies the role of ethanol and ethanol-derived key metabolites [i.e., acetaldehyde (Acet) and phosphatidylethanol (PEth)] will be established. Four specific aims will test this hypothesis using rat hepatocytes and in vivo ethanol-fed rats as models. Aim I will determine the comparative effects of ethanol, Acet and PEth on activation/phosphorylation of p38 MAPK, MSK-1, JNK kinases. In Aim II, effects of ethanol and Acet on nuclear p38 MAPK, MSK-1 and histone-3 phosphorylation will be elucidated using pharmacological inhibitors and antisense strategy and will be correlated to apoptosis. Aim III will determine the "phospho-proteomic map" of nuclear targets affected by the above kinases through functional proteomic technique. These in vitro studies will be extended to in vivo studies in Aim IV, where chronically ethanol fed rat hepatocytes will be used to determine the modulation of p38 MAPK, MSK-1 and histone-3 phosphorylation by ethanol and Acet. The reversibility of the modulatory effects will be also investigated after ethanol withdrawal. Our focus onto nuclear effects of ethanol and its metabolites on specific members of MAPK family highlights a new dimension in ethanol research and will offer novel insight into the molecular mechanisms of ethanol actions at the subcellular organelle level. Histone modifications by ethanol will have mechanistic involvement in chromatin remodeling/gene expression. Relevance of these observations to apoptotic changes will also be established. New knowledge gained from this project will facilitate development of therapeutic tools to prevent and control ethanol-induced cellular damage.

描述（由申请人提供）：乙醇的分子作用仍然很大程度上未知。最近的进展表明，乙醇会改变细胞内信号传导途径，包括涉及丝裂原激活蛋白 (MAP) 激酶家族成员的信号传导途径。基于新的支持数据，假设“乙醇调节 p38 MAPK 活性，导致核丝裂原和应激激活激酶 1 (MSK-1) 的下游激活以及作为靶点的组蛋白 3 的磷酸化。”这一更新应用的一个重要焦点是乙醇对细胞核信号传导及其后果的影响，例如对组蛋白修饰和细胞凋亡的影响。在这些研究中，将确定乙醇和乙醇衍生的关键代谢物 [即乙醛 (Acet) 和磷脂酰乙醇 (PEth)] 的作用。四个具体目标将使用大鼠肝细胞和体内乙醇喂养的大鼠作为模型来检验这一假设。目标 I 将确定乙醇、Acet 和 PEth 对 p38 MAPK、MSK-1、JNK 激酶激活/磷酸化的比较影响。在目标 II 中，将使用药理学抑制剂和反义策略阐明乙醇和 Acet 对核 p38 MAPK、MSK-1 和组蛋白 3 磷酸化的影响，并将与细胞凋亡相关。 Aim III将通过功能蛋白质组技术确定受上述激酶影响的核靶点的“磷酸化蛋白质组图谱”。这些体外研究将扩展到 Aim IV 的体内研究，其中长期乙醇喂养的大鼠肝细胞将用于确定乙醇和 Acet 对 p38 MAPK、MSK-1 和组蛋白 3 磷酸化的调节。乙醇戒断后调节作用的可逆性也将被研究。我们对乙醇及其代谢物对 MAPK 家族特定成员的核效应的关注突出了乙醇研究的新维度，并将为亚细胞细胞器水平上乙醇作用的分子机制提供新的见解。乙醇对组蛋白的修饰将在机制上参与染色质重塑/基因表达。还将确定这些观察结果与细胞凋亡变化的相关性。从该项目中获得的新知识将有助于开发预防和控制乙醇引起的细胞损伤的治疗工具。