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標題: | 尋找輔助性熱休克蛋白Ids2所調控的潛在生理途經 Defining Ids2 co-chaperone controlling pathways |
作者: | Yu-Tung Lo 羅予彤 |
指導教授: | 鄧述諄 |
關鍵字: | 老化,熱休克蛋白,輔助性熱休克蛋白,基因交互作用,合成致死,粒線體功能異常, caloric restriction,proteostasis,chaperone,co-chaperone,synthetic genetic array (SGA),synthetic lethality/sickness, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 老化是生物體逐漸喪失正常生理功能所造成的現象,在過去許多研究中指出營養失調與細胞內蛋白質失衡等因素都是引起老化發生的重要關鍵。其中熱休克蛋白便是調控細胞內蛋白質恆定的主要分子,熱休克蛋白90 (HSP90)是真核生物中被高度保留下的分子,細胞中許多蛋白質的折疊與活性的成熟與否都需要藉由熱休克蛋白90 (HSP90)的調控才能夠正常發揮作用,因此熱休克蛋白90 (HSP90)在許多生理反應途徑中扮演著重要的角色,然而,為了讓熱休克蛋白90 (HSP90)能有效率的運作,過程中需要大量輔助性熱休克蛋白(co-chaperone)的幫忙,這些蛋白質會參與調控熱休克蛋白90 (HSP90)的結構、活性以及對受質(substrate)的專一性。在近期本實驗室所發表的研究中,發現Ids2可能作為調控熱休克蛋白90 (HSP90)的輔助性熱休克蛋白,並且發現在其第148個胺基酸上的磷酸化會妨礙Ids2與熱休克蛋白90 (HSP90)的交互作用,導致細胞內不正常的蛋白質堆積與粒線體功能的異常。然而,當初在不同酵母菌菌株中建構IDS2缺失時,發現在不同菌株中,IDS2缺失會使不同菌株在相同壓力下表現出不同的生長狀況,這使我們思考是否在不同菌株中,某些基因的表現量不相同並且Ids2與這些基因的蛋白質產物可能共同參與在某些生理途徑中,於是我們利用大規模基因交互作用(genetic interaction)的分析找出四個與IDS2有合成致死(synthetic lethality)的基因,這些基因分別為GIM4、EAF1、SKI8與YGR035C,並期望利用後續研究能找出Ids2是如何共同參與在相關的生理途徑中,進而影響細胞對壓力的調控。 In a paper recently published by our lab, we performed mass spectrometry-based phosphoproteomic screening and found that Ids2 (IME2-dependent signaling protein) was dephosphorylated at residue 148 under caloric restriction (CR). Moreover, the phosphorylation /dephosphorylation manner affected the interaction between Ids2 and heat shock protein 90 (HSP90), a highly conserved molecular chaperone in eukaryotes and is necessary for protein folding, maturation and activity of its substrates (which are also termed clients). Chaperones play a critical role in proteostasis which is regarded as one of the hallmarks of aging and regulates many biological processes, such as signal transduction and stress response. To function effectively, HSP90 requires a group of proteins, co-chaperones, to regulate the conformational change, client specificity and activity of HSP90. Based on our previous results, Ids2 may act as a co-chaperon regulating HSP90 activity and the phosphorylation of Ids2 disrupts its interaction with HSP90 leading to abnormal protein aggregation and mitochondrial defect. When we constructed IDS2 deletion in different yeast strains, W303 and BY4741 background strains, we found that the W303 ids2strain exhibited severe growth defect under 37°C heat shock and glycerol-treated medium, while the BY4741 ids2 strain did not. These consequences made us wonder that there might be some difference of gene expression between W303 and BY4741 background strains. We conducted the synthetic genetic array (SGA) analysis and further identified four genes, GIM4, EAF1, SKI8 and YGR035C, that are synthetic lethality/sickness with IDS2. These results suggested that Ids2 might control the pathways with these four genes in some degree and regulate cell viability under different stress conditions. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74867 |
DOI: | 10.6342/NTU201904223 |
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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