CHARACTERIZATION OF NAG EXPRESSION AND FUNCTIONS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 白细胞介素-1受体拮抗剂（IL-1 Ra）是天然存在的白细胞介素-1（IL-1）抑制剂。 IL-1 Ra的两种细胞内形式（icIL-1 Ra 1和icIL-1 Ra 3）已在蛋白质水平上得到描述。 这些异构体的确切功能尚不清楚。 研究已经证明icIL-1 Ra 1作为IL-1诱导的p38 MAP激酶活性、NF κ B易位以及上皮细胞中IL-6和IL-8表达的抑制剂的作用。 先前从酵母双杂交分析获得的数据表明，icIL-1 Ra 1与两种细胞内蛋白质，COP 9信号体亚基3（CSN 3）和神经母细胞瘤扩增基因产物（NAG）相互作用。 CSN 3是COP 9信号体的组分，其与icIL-1 Ra 1的相互作用导致与信号体复合物相关的CK 2和PKD激酶的抑制。 NAG在高级别神经母细胞瘤中与MYCN沿着共扩增，然而NAG蛋白的正常功能尚不清楚。 我们推测icIL-1 Ra 1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，我们启动了实验，检查icIL-1 Ra 1和NAG在相关细胞系统中的相互作用。 分子工具的产生，使我们能够重新确认NAG和icIL-1 Ra 1在相关细胞系统中的相互作用。已经设计了过表达和敲低表达系统，并用于检测干扰NAG和icIL-1 Ra 1的表达对全局基因表达的影响。 通过微阵列分析鉴定的基因表达变化有助于鉴定将NAG和icIL-1 Ra 1与常见生理过程联系起来的候选途径。 目前正在研究这种联系的相关性。

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 under study.