Characterize Bcl-2 Family Function/Regulation

表征 Bcl-2 家族功能/调节

• 批准号: 6867369
• 负责人: SHINE S TU
• 金额: $ 2.57万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2005-08-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6867369
• 关键词: BCL2 gene /protein; DNA damage; SDS polyacrylamide gel electrophoresis; apoptosis; cell line; cytochrome c; cytoskeleton; endoplasmic reticulum; gene targeting; genetic regulation; genetic transcription; genetically modified animals; laboratory mouse; membrane permeability; mitochondrial membrane; p53 gene /protein; polymerase chain reaction; postdoctoral investigator; protein protein interaction; protein structure function; recombinant proteins; transfection; ultraviolet radiation

DESCRIPTION (provided by applicant): Apoptosis plays a variety of important roles in the immune system, and it is the goal of this application to elucidate the role of Bcl-2 family members in cell death. A critical decision point in the regulation of apoptosis occurs when the mitochondrial outer membrane permeabilizes to release proteins from the intermembrane space. The Bcl-2 family of proteins are thought to play a key role in the regulation of mitochondrial membrane permeabilization. Three fundamental aspects of the regulation of the Bcl-2 family of proteins will be addressed. In the first aim, we will examine the transcriptional regulation of the pro-apoptotic Bcl-2 family members during and after apoptotic stimulation (UV Irradiation, p53-induced, DNA damage, ER stress, activation induced cell death, and cytoskeletal disruption). This will be achieved through the use of RT real-time PCR techniques that have been established in the laboratory. In the second aim, we will examine the activation state of pro-apoptotic Bcl-2 family members in response to various apoptotic stimuli. This will be achieved through the use of a novel "Bcl-xL trap" that will interact with activated pro-apoptotic Bcl-2 family members. In the third aim, we will extend our studies to determine which pro-apoptotic Bcl-2 family members are necessary, sufficient, or redundant for induction of apoptosis. This will be achieved through the use of BiRNA. Additionally, we will use a call free apoptotic assay to determine which pro-apoptotic Bcl-2 family members have the ability to induce the release of cytochrome-c. The research outlined in this application focuses on the interaction between the pro-apoptotic and anti-apoptotic Bcl-2 family members in response to apoptotic stimulation. Our studies will elucidate those Bcl-2 family members that are: 1) expressed and/or upregulated, 2) activated, 3) necessary, sufficient, or redundant for induction of apoptosis.

描述（由申请人提供）：细胞凋亡在免疫系统中发挥多种重要作用，本申请的目的是阐明Bcl-2家族成员在细胞死亡中的作用。当线粒体外膜透化以从膜间隙释放蛋白质时，细胞凋亡调节中的关键决策点发生。 Bcl-2 蛋白家族被认为在线粒体膜透化的调节中发挥着关键作用。将讨论 Bcl-2 蛋白家族调节的三个基本方面。第一个目标是，我们将检查促凋亡 Bcl-2 家族成员在凋亡刺激期间和之后的转录调控（紫外线照射、p53 诱导、DNA 损伤、内质网应激、激活诱导的细胞死亡和细胞骨架破坏）。这将通过使用实验室已建立的 RT 实时 PCR 技术来实现。在第二个目标中，我们将检查促凋亡 Bcl-2 家族成员响应各种凋亡刺激的激活状态。这将通过使用新型“Bcl-xL 陷阱”来实现，该陷阱将与激活的促凋亡 Bcl-2 家族成员相互作用。第三个目标是，我们将扩展我们的研究，以确定哪些促凋亡 Bcl-2 家族成员对于诱导细胞凋亡是必要的、充分的或多余的。这将通过使用 BiRNA 来实现。此外，我们将使用无调用细胞凋亡测定来确定哪些促凋亡 Bcl-2 家族成员具有诱导细胞色素 c 释放的能力。本申请中概述的研究重点是促凋亡和抗凋亡 Bcl-2 家族成员之间响应凋亡刺激的相互作用。我们的研究将阐明那些 Bcl-2 家族成员：1）表达和/或上调，2）激活，3）诱导细胞凋亡所必需的、充分的或冗余的。