FUNCTIONAL ANALYSIS OF ANTIAPOPTOTIC IAP PROTEINS

抗凋亡 IAP 蛋白的功能分析

基本信息

批准号：
2681261
负责人：
ROLLIE J CLEM
金额：
$ 9.73万
依托单位：
KANSAS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-07-01 至 2003-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2681261
关键词：
Baculoviridae Lepidoptera animal tissue apoptosis atomic absorption spectrometry cell cycle proteins cell growth regulation chimeric proteins gene expression genetic library immunoprecipitation molecular cloning polymerase chain reaction protein binding protein structure function site directed mutagenesis western blottings yeast two hybrid system

项目摘要

The process of apoptosis, or programmed cell death, is of vital importance in human health and disease. Improved understanding of the regulation of apoptosis promises to lead to new therapeutic strategies for a number of diseases, including cancer, neurological degenerative conditions, and autoimmune dysfunction. Much of our current understanding of the regulation of apoptosis rests on a foundation established by studies in lower animals, including nematodes and insects. An example of the importance of studying cell death in lower animals is provided by the anti-apoptotic gene family called iap. The first iap genes were identified by the PI in baculoviruses, which are viruses of lepidopteran insects. Subsequently, iap homologs have been identified by sequence homology in the genomes of insects and higher animals, and there is evidence linking at least two iap genes to human diseases, namely cancer and spinal muscular atrophy. A novel sequence motif found only in IAP proteins, the BIR motif, is required for anti- death function, presumably through the ability of different BIR motifs to bind to a number of distinct pro-apoptotic proteins. However, the BIR motifs from different IAP proteins do not appear to be inter- changeable, and no detailed information is currently available on the sequence requirements for BIR function. The first two Specific Aims of this proposal address the molecular basis for IAP function, through detailed structure-function analyses and characterization of protein- protein interactions. The third Specific Aim utilizes an improved version of the functional assay used to identify the first iap genes. This improved assay will be used to screen both viral and cellular cDNA libraries for novel anti-apoptotic genes. The identification and characterization of new genes involved in the regulation of cell death will lead to improved understanding of apoptotic pathways in both insects and higher animals, and may also aid in understanding the function of IAP proteins.

细胞凋亡或程序性细胞死亡的过程至关重要对人类健康和疾病的重要性。增进了对细胞凋亡的调节有望带来新的治疗策略治疗多种疾病，包括癌症、神经退行性疾病条件和自身免疫功能障碍。我们现在的很多对细胞凋亡调节的理解有一个基础通过对低等动物（包括线虫和昆虫。研究低等细胞死亡的重要性的一个例子动物体内的抗凋亡基因家族称为iap。这第一个 iap 基因是由 PI 在杆状病毒中鉴定出来的，它们是鳞翅目昆虫的病毒。随后，iap 同系物被通过昆虫及高等动物基因组中的序列同源性来鉴定动物，并且有证据表明至少有两个 iap 基因与人类有关疾病，即癌症和脊髓性肌萎缩症。新颖的序列仅在 IAP 蛋白中发现的基序（BIR 基序）是抗- 死亡功能，大概是通过不同 BIR 基序的能力与许多不同的促凋亡蛋白结合。然而，来自不同 IAP 蛋白的 BIR 基序似乎并不相互关联可能会发生变化，目前还没有详细信息 BIR 功能的序列要求。前两个具体目标该提案通过以下方式解决了 IAP 功能的分子基础详细的结构功能分析和蛋白质表征蛋白质相互作用。第三个具体目标采用了改进的用于鉴定第一个 iap 基因的功能测定的版本。这种改进的检测方法将用于筛选病毒和细胞 cDNA 新型抗凋亡基因文库。身份识别和参与细胞死亡调节的新基因的表征将提高对细胞凋亡途径的了解昆虫和高等动物，也可能有助于理解 IAP 蛋白的功能。