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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49702
標題: | 探討熱量攝取和PKA所調控的熱休克輔助性蛋白磷酸化與細胞壽命的關聯性 Calorie-driven PKA-mediated Co-chaperone Phosphorylation Controls Life Span |
作者: | Hsuan-Ming Chen 陳宣銘 |
指導教授: | 鄧述諄(Shu-Chun Teng) |
關鍵字: | 老化,熱量限制,磷酸化,PKA,Hsp90, aging,calorie restriction,phosphorylation,PKA,HSP90, |
出版年 : | 2016 |
學位: | 碩士 |
摘要: | 老化是種具有生理功能的逐漸喪失相關聯的複雜的現象。其本身是由內在因素和外在因素所調節。以前的研究表明,熱量限制延長壽命主要是透許多磷酸化激酶的參與下,以磷酸化蛋白間的訊息傳遞所調控的.;然而,這些信號的磷酸化途徑是如何協調的詳細分子機制仍然知之甚少。在這裡,我們實驗室利用質譜儀的技術大規模地去尋找在限制熱量攝入的酵母磷酸化和去磷酸化位點。質譜總共定量出1244磷酸化/去磷酸化位點,共632磷酸化蛋白有受到調控的變化。 從發現的磷酸化蛋白選出110個進行功能篩選,最後篩篩到一個受到PKA磷酸化的有絲分裂相關的未知功能蛋白IDS2,在老化進程作出貢獻的一個關鍵調控。 IDS2會扮演著一個輔助性熱休克蛋白,與熱休克蛋白Hsc82和冗餘的Hsp82形成複合物。 IDS2剔除基因或IDS2模擬磷酸化菌株皆會產生熱敏性和壽命縮短的現象。而需要HSP90輔佐其折疊的蛋白會在IDS2剔除或IDS2模擬磷酸化的細胞不穩定。IDS2去磷酸化則是會恢復這些表型。從這些結果皆可得知,酵母菌偵測熱量攝入的變化,進而透過PKA的活化與否去影響到HSP90伴侶相關的折疊的活性,並由此影響個體的壽命。 Aging, an intricate phenomenon associated with the gradual loss of physiological functions, is regulated by intrinsic and extrinsic factors. Previous studies indicate that calorie restriction extends the life span of a variety of species due to the involvement of multiple kinase regulations; however, the molecular mechanism of how these signaling phosphorylation pathways are coordinated is still poorly understood. Here we globally identify yeast phosphorylation and dephosphorylation sites under calorie restriction. Quantitative mass spectrometry identified a total of 1244 phosphorylation/ dephosphorylation sites on 632 proteins. Functional screen of the 110 potential regulators discovered a mitotically functional unknown protein Ids2 phosphorylated by PKA under rich medium, contributing to a key regulation in aging process. Ids2 serves as a cochaperone to form a complex with Hsc82 and the redundant Hsp82. ids2 or ids2 phosphor-mimicking cells displayed heat sensitivity and life span shortening. The in-vivo substrate of HSP90 is destabilized in ids2 or ids2 phosphor-mimicking cells. Dephosphorylation of this Ids2 phosphorylation recovered these phenotypes. Taken together, yeast cells sense calorie intake and compromise HSP90 chaperone folding activity through PKA, and thereby influence the lifespan. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/49702 |
DOI: | 10.6342/NTU201602696 |
全文授權: | 有償授權 |
顯示於系所單位: | 微生物學科所 |
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