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  1. NTU Theses and Dissertations Repository
  2. 醫學院
  3. 藥理學科所
Please use this identifier to cite or link to this item: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26496
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???org.dspace.app.webui.jsptag.ItemTag.dcfield???ValueLanguage
dc.contributor.advisor林琬琬(Wan-Wan Lin)
dc.contributor.authorYu-Cheng Tuen
dc.contributor.author涂育誠zh_TW
dc.date.accessioned2021-06-08T07:12:31Z-
dc.date.copyright2011-10-07
dc.date.issued2011
dc.date.submitted2011-08-11
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26496-
dc.description.abstractPMA 為一已知的蛋白磷酸激酶C (PKC) 活化劑, 可活化轉錄因子NF-kappaB及c-Fos蛋白表現。我們欲探討此兩條活化路徑的關係,因此比較腫瘤壞死因子-alpha (TNF-alpha) 和PMA在不同種細胞的作用。首先,我們發現PMA所誘導的p65蛋白絲胺酸 (Serine) 536磷酸化需要IKKalpha及IKKbeta,而TNF-alpha的作用則非如此。除此之外,在多種細胞我們發現TNF-alpha 會引起強烈IKKalpha/beta 活性區的絲胺酸176及絲胺酸177磷酸化,相反的,PMA偏好於IKKalpha/beta活性區的絲胺酸180及絲胺酸181產生磷酸化,並且此磷酸化作用與PKC-MEK-ERK路徑有關,但與RSK及MSK無關。然而,此種磷酸化作用並未參與PMA所誘導的IkappaBalpha蛋白降解、p65蛋白絲胺酸536磷酸化、p65蛋白進入細胞核以及轉錄因子NF-kappaB的活化。出乎意料,我們發現PMA所誘導的c-Fos蛋白表現需要IKKalpha/IKKbeta/IKKgamma複合物,而此種相依性也在p65蛋白缺陷的老鼠胚胎纖維母細胞 (MEF) 中發現。更進一步,我們在老鼠c-fos基因啟動子 (promoter) 區域中找到了一個可能被轉錄因子NF-kappaB所結合的區域。經由實驗證明,在PMA的刺激之下,p65蛋白同質雙聚體 (p65 homodimer) 可以和CBP/p300蛋白結合到此區域進行基因調控,而此種調控與TCF-SRF-SRE複合物相關之調控無關。簡而言之,此篇研究發現IKKalpha/beta蛋白磷酸化的新機制,也證實PMA所誘導的c-fos基因調控需要IKKalpha/beta蛋白參與,而c-fos基因啟動子之轉錄因子NF-kappaB結合位置的發現,將可對於轉錄因子NF-kappaB及轉錄因子AP-1兩者之間的關係,提供一個新的思考點及研究方針。zh_TW
dc.description.abstractPhorbol 12-myristate 13-acetate (PMA) is a PKC activator known for triggering NF-kappaB activation and c-fos expression. To further understand the functional link of both events, we compared the actions of TNF-alpha and PMA in various cell types. We found that PMA- but not TNF-alpha-induced p65 Ser536 phosphorylation in mouse embryonic fibroblast (MEF) is IKKalpha/beta dependent. Additionally, compared to TNF-alpha, which induced dramatically IKKalpha/beta phosphorylation at Ser176/177, PMA preferentially induced IKKalpha/beta phosphorylation at Ser180/181 within its active loop, and this phosphorylation of IKKalpha/beta by PMA is through a novel mechanism depending on PKC-MEK-ERK axis but not RSK or MSK. Moreover, this phosphorylation does not contribute to PMA-induced IkappaBalpha degradation, p65 Ser536 phosphorylation, p65 nuclear translocation and NF-kappaB activation. Surprisingly, we found that PMA-induced immediately early gene c-fos expression is suppressed in IKKalpha-/-, IKKbeta-/-, IKKgamma-/- and p65-/- MEF, suggesting the involvement of NF-kappaB in c-fos gene transcription. Furthermore, we indentified a previously uncharacterized NF-kappaB binding site located in the mouse c-fos promoter. Under PMA stimulation, p65 homodimer in association with CBP/p300 is capable of binding to this site, and this action is independent of TCF-SRF-SRE ternary complex regulation. In summary, this study provides different views of IKK phosphorylation mechanism. Importantly, the requirement of IKK complex in PMA-induced c-fos expression and the NF-kappaB binding site found in c-fos promoter shed light on the crosstalk between NF-kappaB and AP-1 transcription factors.en
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Previous issue date: 2011
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dc.description.tableofcontentsAbbreviations.…………………………………………………………I
Abstract…………………………………………………………………VI
中文摘要…………………………………………………………………IV
Introduction……………………………………………………………1
Materials and Methods………………………………………………23
Results…………………………………………………………………35
Discussion……………………………………………………………51
Figures…………………………………………………………………65
Appendix………………………………………………………………90
References……………………………………………………………105
dc.language.isoen
dc.subject啟動子zh_TW
dc.subject磷酸化zh_TW
dc.subject絲胺酸zh_TW
dc.subject轉錄因子zh_TW
dc.subjectNF-kappaBen
dc.subjectc-fosen
dc.subjectPMAen
dc.subjectIKKen
dc.title探討IKK調控PMA誘導c-fos基因表現之機轉zh_TW
dc.titleStudy on the IKK-dependent regulation of PMA-induced c-fos gene expressionen
dc.typeThesis
dc.date.schoolyear99-2
dc.description.degree碩士
dc.contributor.oralexamcommittee符文美(Wen-Mei Fu),蘇銘嘉(Ming-Jai Su),徐立中(Li-Chung Hsu),曾炳輝(Ping-Hui Tseng)
dc.subject.keyword轉錄因子,磷酸化,啟動子,絲胺酸,zh_TW
dc.subject.keywordIKK,NF-kappaB,PMA,c-fos,en
dc.relation.page128
dc.rights.note未授權
dc.date.accepted2011-08-11
dc.contributor.author-college醫學院zh_TW
dc.contributor.author-dept藥理學研究所zh_TW
Appears in Collections:藥理學科所

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