Oh-hashi Kentaro, Koga Hisashi, Ikeda Shun, Shimada Kiyo, Hirata Yoko, Kiuchi Kazutoshi
Department of Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu, Japan.
Biochem Biophys Res Commun. 2009 Sep 25;387(3):504-10. doi: 10.1016/j.bbrc.2009.07.047. Epub 2009 Jul 15.
Recently, endoplasmic reticulum (ER) stress responses have been suggested to play important roles in maintaining various cellular functions and to underlie many tissue dysfunctions. In this study, we first identified cysteine-rich with EGF-like domains 2 (CRELD2) as an ER stress-inducible gene by analyzing a microarray analysis of thapsigargin (Tg)-inducible genes in Neuro2a cells. CRELD2 mRNA is also shown to be immediately induced by treatment with the ER stress-inducing reagents tunicamycin and brefeldin A. In the genomic sequence of the mouse CRELD2 promoter, we found a typical ER stress responsible element (ERSE), which is well conserved among various species. Using a luciferase reporter analyses, we demonstrated that the ERSE in mouse CRELD2 is functional and responds to Tg and ATF6-overexpression. Each mutation of ATF6- or NF-Y-binding sites in the ERSE of the mouse CRELD2 promoter dramatically decreased both the basal activity and responsiveness toward the ER stress stimuli. Our study suggests that CRELD2 could be a novel mediator in regulating the onset and progression of various ER stress-associated diseases.
最近，内质网（ER）应激反应被认为在维持各种细胞功能中发挥重要作用，并且是许多组织功能障碍的基础。在本研究中，我们首先通过分析毒胡萝卜素（Tg）诱导Neuro2a细胞基因的微阵列分析，确定富含半胱氨酸且具有表皮生长因子样结构域2（CRELD2）为内质网应激诱导基因。CRELD2 mRNA也被证明在用内质网应激诱导试剂衣霉素和布雷菲德菌素A处理后会立即被诱导。在小鼠CRELD2启动子的基因组序列中，我们发现了一个典型的内质网应激反应元件（ERSE），它在各种物种中高度保守。通过荧光素酶报告基因分析，我们证明小鼠CRELD2中的ERSE具有功能，并且对Tg和ATF6过表达有反应。小鼠CRELD2启动子的ERSE中ATF6或NF-Y结合位点的每个突变都显著降低了基础活性和对内质网应激刺激的反应性。我们的研究表明，CRELD2可能是调节各种内质网应激相关疾病发生和发展的新型介质。
