ANALYSIS OF ICIL-1RA1 AND NAG INTERACTIONS

ICIL-1RA1 和 NAG 相互作用的分析

• 批准号: 7381640
• 负责人: CARLA J GUTHRIDGE
• 金额: $ 11.14万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381640
• 关键词: ANALYSIS; ICIL; 1RA1; NAG; INTERACTIONS

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Interleukin-1 receptor antagonists (IL-1Ra) are naturally occurring inhibitors of interleukin-1 (IL-1). Two intracellular forms of IL-1Ra (icIL-1Ra1 and icIL-1Ra3) have been described at the protein level. The precise function of these isoforms is unclear. Studies have demonstrated a role for icIL-1Ra1 as an inhibitor of IL-1-induced p38 MAP kinase activity, NFkB translocation, and IL-6 and IL-8 expression in epithelial cells. Previous data obtained from yeast two-hybrid analysis suggested that icIL-1Ra1 interacts with two intracellular proteins, COP9 signalosome subunit 3 (CSN3) and neuroblastoma amplified gene product (NAG). CSN3 is a component of the COP9 signalosome and its interaction with icIL-1Ra1 leads to an inhibition of CK2 and PKD kinases that associate with the signalosome complex. NAG is co-amplified along with MYCN in high grade neuroblastomas, however the normal function of the NAG protein is unknown. We hypothesize that icIL-1Ra1 and NAG play a crucial role in the regulation of cellular processes associated with IL-1 signaling. To address this hypothesis we initiated experiments examining the interaction of icIL-1Ra1 and NAG in relevant cell systems. Molecular tools were generated that allowed us to reconfirm the interaction of NAG and icIL-1Ra1 in relevant cell systems. Over-expression and knock-down expression systems have been designed and used to examine the effect of perturbing the expression of NAG and icIL-1Ra1 on global gene expression. Gene expression changes identified by microarray analysis have helped to identify candidate pathways linking NAG and icIL-1Ra1 to common physiological processes. The relevance of this linkage is currently unde

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。白介素-1受体拮抗剂（IL-1RA）是白介素-1（IL-1）的天然抑制剂。在蛋白质水平上描述了两种细胞内IL-1RA（ICIL-1RA1和ICIL-1RA3）。这些同工型的确切功能尚不清楚。研究表明，ICIL-1RA1作为IL-1诱导的p38 MAP激酶活性，NFKB易位以及IL-6和IL-8表达在上皮细胞中的作用。从酵母双杂交分析获得的先前数据表明，ICIL-1RA1与两种细胞内蛋白相互作用，COP9信号亚基3（CSN3）和神经母细胞瘤扩增基因产物（NAG）。 CSN3是COP9信号体的一个组成部分，其与ICIL-1RA1的相互作用会导致抑制与信号体复合物相关的CK2和PKD激酶。 NAG与MYCN一起在高级神经母细胞瘤中共同扩增，但是NAG蛋白的正常功能尚不清楚。我们假设ICIL-1RA1和NAG在调节与IL-1信号相关的细胞过程中起着至关重要的作用。为了解决这一假设，我们启动了检查相关细胞系统中ICIL-1RA1和NAG相互作用的实验。生成了分子工具，使我们能够在相关细胞系统中重新确认NAG和ICIL-1RA1的相互作用。过表达和敲门表达系统已被设计和用于检查NAG和ICIL-1RA1表达对全局基因表达的表达的影响。通过微阵列分析确定的基因表达变化有助于确定将NAG和ICIL-1RA1与常见生理过程联系起来的候选途径。此链接的相关性目前是UNDE