Yaykasli Kursat Oguz, Oohashi Toshitaka, Hirohata Satoshi, Hatipoglu Omer Faruk, Inagawa Kiichi, Demircan Kadir, Ninomiya Yoshifumi
Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan.
Mol Cell Biochem. 2009 Mar;323(1-2):69-79. doi: 10.1007/s11010-008-9965-4. Epub 2008 Dec 4.
ADAMTS9 is a member of the disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) genes, with aggrecan-degrading activity. It has also been characterized to be reactive and highly activated ADAMTS by IL-1 beta in both chondrosarcoma cells and human chondrocytes (Demircan et al. Arthritis Rheum 52:1451-1460, 2005). In order to understand the regulation of ADAMTS9 gene expression a functional 3.0 kb human ADAMTS9 promoter has been cloned and characterized. A sequence analysis of the promoter revealed the presence of putative binding sites for Nuclear Factor of Activated T cells (NFAT), which is commonly found in the ADAMTS4 and ADAMTS5 promoters. NFATc1 was up-regulated in an activated form by IL-1 beta in human chondrocytes. The IL-1 beta inducible ADAMTS9 expression was inhibited by NFAT inhibitors, FK506 and 11Arg (11R)-VIVIT. Furthermore, direct binding of NFATc1 on distal and proximal promoters of ADAMTS9 was demonstrated by a chromatin immunoprecipitation assay. Promoter-reporter assays supported those results. These findings may provide a better understanding of the regulation of ADAMTS9 expression induced by inflammatory cytokines.
ADAMTS9是具有血小板反应蛋白基序的解聚素和金属蛋白酶（ADAMTS）基因家族的成员，具有蛋白聚糖降解活性。在软骨肉瘤细胞和人软骨细胞中，它还被证明对白细胞介素-1β有反应且高度活化（Demircan等人，《关节炎与风湿病》52:1451 - 1460，2005年）。为了了解ADAMTS9基因表达的调控机制，已克隆并鉴定了一个3.0 kb的人ADAMTS9功能性启动子。对该启动子的序列分析揭示了活化T细胞核因子（NFAT）的假定结合位点的存在，这在ADAMTS4和ADAMTS5启动子中很常见。在人软骨细胞中，白细胞介素-1β以活化形式上调NFATc1。NFAT抑制剂FK506和11Arg（11R）-VIVIT抑制了白细胞介素-1β诱导的ADAMTS9表达。此外，染色质免疫沉淀试验证明了NFATc1与ADAMTS9远端和近端启动子的直接结合。启动子报告基因试验支持了这些结果。这些发现可能有助于更好地理解炎性细胞因子诱导的ADAMTS9表达的调控机制。
