本文關(guān)鍵詞：熱休克蛋白90與TAK1相互作用調(diào)節(jié)白介素-1β激活的信號轉(zhuǎn)導　出處：《南京師范大學》2008年碩士論文　論文類型：學位論文

  更多相關(guān)文章： 熱休克蛋白90 轉(zhuǎn)化生長因子-β活化激酶1 格爾德霉素 白介素-1β 炎癥 信號轉(zhuǎn)導

【摘要】： 熱休克蛋白90(Heat shock protein 90,Hsp90)是Hsp90家族成員,進化上高度保守,其結(jié)構(gòu)主要由一個N端高度保守的ATPase結(jié)構(gòu)域,一個中間結(jié)構(gòu)域以及一個介導Hsp90二聚化的C端結(jié)構(gòu)域三部分組成。Hsp90是胞內(nèi)廣泛存在的一個分子伴侶,其底物蛋白主要為一些在細胞內(nèi)信號通路中起重要作用的類固醇激素受體、蛋白激酶、轉(zhuǎn)錄因子等。Hsp90特異性抑制劑GA或RAD,能夠通過競爭的結(jié)合到Hsp90 N端的ATP/ADP結(jié)合位點阻斷其分子伴侶功能,從而導致其底物蛋白通過蛋白酶體降解。因此,借助于Hsp90的抑制劑可以進一步擴大Hsp90的底物范圍。IL-1是一種促炎細胞因子,主要由LPS刺激的單核/巨嗜細胞產(chǎn)生,在機體免疫反應和免疫缺陷性疾病如類風濕關(guān)節(jié)炎(rheumatoid arthritis,RA)中具有重要作用。TAK1作為MAPKKK參與IL-1β誘導的信號通路,并對下游MAPKs(JNK和p38)和NF-κ3的激活是必需的。蛋白質(zhì)組學研究表明Hsp90能夠與TAK1相互作用。 我們的研究發(fā)現(xiàn)并報道了Hsp90在IL-1β激活的信號級聯(lián)中的新的調(diào)控作用。GA處理后抑制了IL-1β誘導的,TAK1-MAPKs和TAK1-NF-κ3通路的激活,從而導致炎癥相關(guān)蛋白COX-2表達的減少。利用RNA干擾手段下調(diào)內(nèi)源Hsp90的蛋白水平后,我們也得到了相同的結(jié)果。此外,T6RZC穩(wěn)定表達細胞系可以通過Coumermycin A誘導T6RZC二聚化,從而模擬IL-1β激活的胞內(nèi)信號級聯(lián),以避開Hsp90分子伴侶復合物對TAK1上游激酶IRAK-1的調(diào)節(jié)作用對MAPKs和NF-κB激活的影響。在T6RZC穩(wěn)定表達細胞系中,GA明顯抑制了JNK、p38 MAPKs的磷酸化和IκB的降解。免疫沉淀實驗結(jié)果顯示Hsp90能夠在體內(nèi)與TAK1發(fā)生相互作用,Hsp90的氨基端1-401位氨基酸介導了兩者的結(jié)合。Hsp90的抑制劑能夠在轉(zhuǎn)錄后水平下調(diào)內(nèi)源TAK1的表達而不影響其mRNA水平,并且GA處理后TAK1蛋白的降解是通過蛋白酶體途徑。這些結(jié)果表明Hsp90通過與TAK1發(fā)生蛋白間相互作用來調(diào)節(jié)IL-1β激活的MAPKs和NF-κB信號轉(zhuǎn)導通路。 綜上所述,本研究首次揭示了Hsp90通過調(diào)節(jié)TAK1蛋白的穩(wěn)定性參與IL-1β激活的信號級聯(lián);提出了Hsp90在胞內(nèi)調(diào)節(jié)MAPKs和NF-κB通路的新的作用機制。這些結(jié)果對于深入研究Hsp90在炎癥相關(guān)信號通路中的調(diào)節(jié)作用具有重要的指導意義。
[Abstract]:Heat shock protein 90 (heat shock protein 90 Hsp90) is a member of the Hsp90 family and is highly conserved in evolution. Its structure is mainly composed of a highly conserved N-terminal ATPase domain. One intermediate domain and one C-terminal domain that mediates Hsp90 dimerization consist of three parts. Hsp90 is a ubiquitous molecular chaperone in the cell. Its substrate proteins are some steroid hormone receptors protein kinases transcription factors and other specific inhibitors GA or RAD which play an important role in intracellular signaling pathway. The molecular chaperone function can be blocked by competitive binding to the ATP/ADP binding site at the N-terminal of Hsp90, which leads to the degradation of its substrate protein through proteasome. The inhibitor of Hsp90 can further expand the range of substrates of Hsp90. IL-1 is a pro-inflammatory cytokine which is mainly produced by mononuclear / giant eosinophil stimulated by LPS. Rheumatoid arthritis in immune response and immunodeficient diseases such as rheumatoid arthritis. TAK1 plays an important role in IL-1 尾 -induced signaling pathway. TAK1 plays an important role in IL-1 尾 -induced signaling pathway. Activation of downstream MAPKs(JNK and p38) and NF- 魏 3 were necessary. Proteomic studies showed that Hsp90 could interact with TAK1. We found and reported the new regulation of Hsp90 in IL-1 尾 -activated signal cascades. Ga treatment inhibited IL-1 尾 -induced signaling. Activation of TAK1-MAPKs and TAK1-NF- 魏 3 pathway. This resulted in a decrease in the expression of inflammatory related protein COX-2. The same results were obtained after RNA interference was used to down-regulate the protein level of endogenous Hsp90. T6RZC stable expression cell line can induce T6RZC dimerization through Coumermycin A, thus mimicking the intracellular signal cascade activated by IL-1 尾. In order to avoid the effect of Hsp90 chaperone complex on the activation of MAPKs and NF- 魏 B in TAK1 upstream kinase IRAK-1, T6RZC stable expression cell line. Ga inhibited the phosphorylation of MAPKs and the degradation of I 魏 B. the immunoprecipitation results showed that Hsp90 could interact with TAK1 in vivo. The amino terminal 1-401 amino acids of Hsp90 mediate that the inhibitor of Hsp90 can down-regulate the expression of endogenous TAK1 at post-transcriptional level without affecting its mRNA level. The degradation of TAK1 protein after GA treatment was mediated by proteasome pathway. These results suggest that Hsp90 regulates the activation of IL-1 尾 -activated MAPKs by interacting with TAK1 proteins. NF- 魏 B signal transduction pathway. In conclusion, this study for the first time revealed that Hsp90 participates in the signal cascade of IL-1 尾 activation by regulating the stability of TAK1 protein. A new mechanism of intracellular regulation of MAPKs and NF- 魏 B pathway by Hsp90 was proposed. These results are important for further study of the regulatory role of Hsp90 in inflammatory signaling pathway. Guide meaning.
【學位授予單位】：南京師范大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R341

【引證文獻】

相關(guān)碩士學位論文 前1條

1 李璐;應用抑制消減雜交技術(shù)（SSH）篩選癲癇的相關(guān)基因[D];山東大學;2012年

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