Redox regulation of Cellular responses against cytokine stress

细胞因子应激反应的氧化还原调节

• 批准号: 09671558
• 负责人: HIROTA Kiichi
• 金额: $ 2.3万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671558/
• 关键词: cytokinemin; redox regulation; thioredoxin; glutaredoxin; transcription factor; Ap-1; NAPK

Oxidative stresses such as ultraviolet (UV) irradiation to mammalian cells triggers variety of oxystress responses including activation of transcription factors. Recently, activation of nuclear factor-kappa B (NF-kappaB) has been shown to be under oxido-reduction (redox) regulation controlled by thioredoxin (TRX), which is one of major endogenous redox-regulating molecules with thiol reducing activity. In order to elucidate where in the cellular compartment TRX participates in NF-kappaB regulation, we investigated the intracellular localization of TRX UVB irradiation induced translocation of TRX from the cytoplasm into the nucleus. In in vitro diamide-induced crosslinking study, we showed that TRX can associate directly with NF-kappaB p50. Overexpression of wild-type TRX suppressed induction of luciferase activity under NF-kappaB-binding sites in response to UV irradiation compared to the mock transfectant. In contrast, overexpression of nuclear-targeted TRX enhanced the luciferase activity. Thus, TRX seems to play dual and opposing roles in the regulation of NF-kappaB, In the cytoplasm, it interferes with the signals to IkappaB kinases and block the degradation of IkappaB.In the nucleus, however, TRX enhances NF-kappaB transcriptional activities by enhancing its ability to bind DNA.This two step TRX-dependent regulation of NP-kappaB complex, may be a novel activation mechanism of redox-sensitive transcription factors.

氧化应激如紫外线（UV）照射到哺乳动物细胞引发各种氧化应激反应，包括转录因子的激活。近年来，核因子κ B （nf - κ B）的活化受到硫氧还蛋白（TRX）的氧化还原（氧化还原）调控，而硫氧还蛋白是主要的内源性氧化还原调节分子之一，具有巯基还原活性。为了阐明TRX在细胞区室中参与NF-kappaB调控的位置，我们研究了UVB照射诱导TRX从细胞质转运到细胞核的细胞内定位。在体外二胺诱导交联研究中，我们发现TRX可以直接与NF-kappaB p50结合。与模拟转染相比，野生型TRX的过表达抑制了紫外线照射下nf - kappab结合位点荧光素酶活性的诱导。相反，核靶向TRX的过表达增强了荧光素酶的活性。因此，TRX似乎在调节NF-kappaB中起着双重和相反的作用，在细胞质中，它干扰IkappaB激酶的信号，阻断IkappaB的降解。然而，在细胞核中，TRX通过增强NF-kappaB结合DNA的能力来增强其转录活性。这种依赖trx的两步调控NP-kappaB复合体，可能是氧化还原敏感转录因子的一种新的激活机制。

项目成果

M,Hotta: "Pancreatic beta cell-specific expression of thioredoxin, an antioxidative and antiapoptotic protein, prevents autoimmune and streptozotocin-induced diabetes." J.Exp.Med. 188. 1445-1451 (1998)

M，Hotta：“硫氧还蛋白（一种抗氧化和抗凋亡蛋白）的胰腺β细胞特异性表达可预防自身免疫和链脲佐菌素诱导的糖尿病。”

Y,Takagi: "Expression of thioredoxin is enhanced in atherosclerotic plaques and during neointima formation in rat" Laboratory Investigation. 78. 957-966 (1998)

Y，Takagi：“硫氧还蛋白的表达在大鼠动脉粥样硬化斑块和新内膜形成过程中增强”实验室研究。

Nishiyama, A., Furuke, K., Hirota, K., Masutani, H., and Yodoi, J.: "Detection of a nuclear 60-kDa protein coimminoprecipitates with human thiordoxin." Oxygen homeostasis and its dynamics ; Keio university synposia for life science and medicine. Vol.1 (Sp

Nishiyama, A.、Furuke, K.、Hirota, K.、Masutani, H. 和 Yodoi, J.：“检测核 60-kDa 蛋白与人硫氧还蛋白共沉淀。”

K,Hirota: "AP-1 Transcriptional Activity is Regulated by a Direct Association between Thioredoxin and Ref-1" Proc. Natl. Acad. Sci. USA. 94. 3633-3638 (1997)

K，Hirota：“AP-1 转录活性由硫氧还蛋白和 Ref-1 之间的直接关联调节”Proc。

Ryuji Yamaguchi, Yuichi Mazaki, Kiichi Hirota, Shigeru Hashimoto, and Hisataka Sabe: "Mitosis specific serine phosphorylation and downregulation of one of the focal adhesion proteins, paxillin." Oncogene. 15. 1753 (1997)

Ryuji Yamaguchi、Yuichi Mazaki、Kiichi Hirota、Shigeru Hashimoto 和 Hisataka Sabe：“有丝分裂特异性丝氨酸磷酸化和粘着斑蛋白之一桩蛋白的下调。”