Pathophysiological mechanism through the regulation of the transcriptional factor, c-fos, gene expression.

病理生理机制通过转录因子c-fos、基因表达的调节。

基本信息

批准号：
11670437
负责人：
IGARASHI Tetsuya
金额：
$ 1.86万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670437/
关键词：
stress stimuli heat shock c-fos prostaglandin endocrine disruptor estrogen 転写調節

项目摘要

1) Elevation of c-fos gene and AP-1 activity induced by stressesAlthough it has been widely accepted that the c-fos gene, which is responsive to various stresses, does not harbor the heat shock element (HSE) in its upstream region, we found the consensus-sequence for the HSE in the human, rat and mouse genes (BBRC 254:566, 1999). Since rostaglandin A1 (PGA1) increases heat shock element (HSE)-mediated transcription, thereby enhancing expression of HSE-bearing genes, including heat shock proteins, we hypothesized that PGA1 might increase c-fos expression through the HSE. Following investigation has revealed that c-fos expression at least partly by increasing the binding between heat shock factor-1 and the HSE, and that PGA1 enhances activity of activating protein-1 (AP-1). Interestingly, so far as PGA1 is present in the medium, AP-1-mediated transcription enhanced by PGA1 cannot be detected by the standard luciferase reporter gene assay. Instead, it can be detected by either checking lu … More ciferase mRNA levels in the presence of PGA1 or measuring luciferase activities just after removal of PGA1. These results shows that protein products of some stress-responsive genes can increase, not during the stressful condition, but immediately after recovery from stress(MCE 164:77, 2000).2) Detection of environmental hormone-like substancesIn the process of the studies on HSE, we have been interested in estrogenic activity because it can enhance expression of c-fos gene as well as heat shock proteins. Since we often experienced troubles with the assay for luciferase in control dishes, we got suspicious of the existence of hormone-like substances with estrogenic activity released from the culture dishes. Our experiments reveals that many types of polystyrene dishes release estrogenic impurities, a property that should be taken into account when carrying out research using estrogen-responsive cells or studying the cellular effects of this hormone and its analogs (Nature Biotechnology 19:812, 2001). Less

1) 应激引起的c-fos基因和AP-1活性升高尽管人们普遍认为，对各种应激有反应的c-fos基因在其上游区域不含有热休克元件(HSE)，我们在人类、大鼠和小鼠基因中发现了 HSE 的共有序列（BBRC 254:566, 1999），因为 rostaglandin A1 (PGA1) 会增加热休克元素。 (HSE) 介导的转录，增强 HSE 承载基因的表达，从而包括热休克蛋白，我们发现 PGA1 可能通过 HSE 增加 c-fos 表达，随后的研究表明，c-fos 表达至少部分是通过增加热休克因子 1 和 HSE 之间的结合，以及 PGA1 增强激活蛋白 1 (AP-1) 的活性意味着，只要培养基中存在 PGA1，PGA1 就不能增强 AP-1 介导的转录。相反，可以通过检查 PGA1 存在下的荧光素酶 mRNA 水平或在去除 PGA1 后测量荧光素酶活性来检测。反应基因的增加不是在压力条件下，而是在压力恢复后立即增加(MCE 164:77, 2000)。2) 环境激素样物质的检测在压力恢复过程中在 HSE 的研究中，我们对雌激素活性很感兴趣，因为它可以增强 c-fos 基因以及热休克蛋白的表达，因为我们在对照培养皿中经常遇到荧光素酶测定的问题，所以我们对激素的存在产生了怀疑。我们的实验表明，许多类型的聚苯乙烯培养皿会释放雌激素杂质，在使用雌激素反应细胞进行研究或研究这种激素的细胞作用时应考虑到这一特性。及其类似物（Nature Biotechnology 19:812, 2001）。