Implication of transcriptional coactivator complexes in rheumatoid synovial cells

转录共激活复合物在类风湿滑膜细胞中的意义

基本信息

批准号：
12557045
负责人：
NAKAJIMA Toshihiro
金额：
$ 6.59万
依托单位：
St. Marianna University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557045/
关键词：
transcriptional coactivator rheumatoid arthritis CREB binding protein (CBP)synovial cell Notch 1 yeast 2 hybrid screening acetylation tumor necrosis factor (TNF)-α 細胞周期 CREB結合蛋白質 CREB結合タンパク質(CBP)Notch-1 腫瘍壊死因子(TNF-α)presenilin

项目摘要

To assess the implication of transcriptional coactivator in rheumatoid synoviocytes activation, we attempted the following two approaches. One is evaluation of nuclear acetylation, one of the critical roles as coactivator function of CREB binding protein (CBP). The other is molecular cloning of CBP-interaction proteins in rheumatoid synoviocytes by yeast two-hybrid systems.1) Using a specific antibody against acetylated lysine, we detected potent nuclear acetylation in rheumatoid synoviocytes and revealed that p53 is one of characteristically acetylated proteins. We further found that TNFα induced acetylation of p53 but attenuated its transcriptional activation in synoviocytes. As overexpression of CBP resulted in p53 promoter activation by TNFα, CBP recruitment is required for p53 activation.2) We cloned Notch-1 as one of CBP-binding proteins from cDNA library prepared from rheumatoid synoviocytes. Notch-1 signal was characteristically activated in rheumatoid synoviocytes though TNFα signaling and implicated in synoviocytes proliferation by TNFα.These results clearly suggested that coactivator takes crucial roles for rheumatoid synoviocytes activation and might contribute to clarifying the pathogenesis of RA.

为了评估转录共激活因子在类风湿性滑膜细胞激活中的作用，我们尝试了以下两种方法：一种是评估核乙酰化，这是 CREB 结合蛋白 (CBP) 共激活因子功能的关键作用之一；另一种是 CBP- 的分子克隆。通过酵母双杂交系统研究类风湿性滑膜细胞中的相互作用蛋白。1）使用针对乙酰化赖氨酸的特异性抗体，我们检测到了强效的核乙酰化我们进一步发现，TNFα 诱导 p53 乙酰化，但减弱了其在滑膜细胞中的转录激活，因为 CBP 的过度表达导致 p53 启动子被 TNFα 激活，因此 p53 激活需要 CBP 的募集。 2)我们从类风湿滑膜细胞制备的cDNA文库中克隆了Notch-1作为CBP结合蛋白之一。 Notch-1 信号在类风湿性滑膜细胞中通过 TNFα 信号传导被特征性激活，并与 TNFα 引起的滑膜细胞增殖有关。这些结果清楚地表明，共激活剂在类风湿性滑膜细胞激活中发挥着至关重要的作用，并可能有助于阐明 RA 的发病机制。