【摘要】： 釀酒酵母ScRck2p是一種MAP Knase激活的蛋白激酶,被Hog1p磷酸化而激活,響應(yīng)于細胞對外界高滲透壓脅迫和氧化脅迫。在本工作中我們發(fā)現(xiàn)釀酒酵母ScRCK2基因的缺失能夠?qū)е录毎麑OR抑制劑——雷帕霉素敏感,但ScRCK2在細胞對雷帕霉素敏感過程中的作用并不依賴于其激酶活性。在人體內(nèi)最常見的機會致病菌-白念珠菌中,我們鑒定了ScRCK2的同源基因CaRCK2,并發(fā)現(xiàn)CaRCK2基因的缺失同樣導(dǎo)致白念珠菌細胞對雷帕霉素敏感,CaRck2p的這一作用同樣不依賴于其激酶活性。此外,我們發(fā)現(xiàn)白念珠菌CaHOG1的缺失也導(dǎo)致細胞對雷帕霉素的敏感。這些結(jié)果表明在釀酒酵母菌和白念珠菌中,作用于HOG途徑下游的RCK2可能調(diào)控TOR信號途徑的功能。 同時,我們發(fā)現(xiàn)白念珠菌CaRCK2基因的缺失能夠?qū)е录毎麑Ω邼B脅迫、氧化脅迫和細胞壁完整性脅迫試劑敏感。比較而言,釀酒酵母ScRCK2基因的缺失只引起細胞對氧化脅迫敏感,而不影響細胞對高滲脅迫和細胞壁完整性脅迫試劑的敏感。但是,釀酒酵母ScRCK2基因卻能夠彌補白念珠菌CaRCK2基因缺失株對高滲脅迫、氧化脅迫和細胞壁完整性脅迫試劑的敏感性表型。這些結(jié)果說明RCK2在細胞內(nèi)發(fā)揮多種功能,但在白念珠菌和釀酒酵母菌中的功能存在分歧。 我們還發(fā)現(xiàn),ScRCK2上游激酶基因ScHOG1和ScPBS2的缺失,以及受ScHOG1調(diào)控的轉(zhuǎn)錄因子基因ScHOT1,ScMSN1,ScMSN2,ScMSN4或ScRLM1的缺失,不影響酵母細胞對雷帕霉素的敏感性。但是,白念珠菌CaHOG1基因的缺失能夠?qū)е录毎麑着撩顾孛舾小＿@些結(jié)果表明白念珠菌HOG途徑與釀酒酵母HOG途徑在細胞功能調(diào)控方面存在分歧,它參與細胞對雷帕霉素敏感性的調(diào)控。
[Abstract]:Saccharomyces cerevisiae ScRck2p (Saccharomyces cerevisiae) is a protein kinase activated by MAP Knase, which is activated by Hog1p phosphorylation. In this work, we found that the absence of ScRCK2 gene in Saccharomyces cerevisiae can cause cells to be sensitive to rapamycin, an inhibitor of TOR, but the role of ScRCK2 in the process of cell sensitivity to rapamycin does not depend on its kinase activity. In Candida albicans, the most common opportunistic pathogen in humans, we identified the homologous gene of ScRCK2, CaRCK2, and found that the deletion of CaRCK2 gene also led to the same effect of Candida albicans cells to rapamycin-sensitive CaRck2P, which was also independent of its kinase activity. In addition, we found that the absence of Candida albicans CaHOG1 also led to cell sensitivity to rapamycin. These results suggest that in Saccharomyces cerevisiae and Candida albicans, RCK2 acting on the downstream of HOG pathway may regulate the function of TOR signaling pathway. At the same time, we found that the deletion of CaRCK2 gene in Candida albicans could result in cell sensitivity to hyperosmotic stress, oxidative stress and cell wall integrity stress. In contrast, the absence of ScRCK2 gene in Saccharomyces cerevisiae only caused the sensitivity of cells to oxidative stress, but not to the sensitivity of cells to hyperosmotic stress and cell wall integrity stress. However, the ScRCK2 gene of Saccharomyces cerevisiae can compensate for the sensitive phenotype of Candida albicans CaRCK2 gene deficient strain to hyperosmotic stress, oxidative stress and cell wall integrity stress. These results suggest that RCK2 plays many functions in cells, but its functions in Candida albicans and Saccharomyces cerevisiae are different. We also found that the deletion of ScHOG1 and ScPBS2 genes upstream of ScRCK2, and the deletion of the transcription factor gene ScHOT1, ScMSN1, ScMSN2, ScMSN2, scMSN4 or ScRLM1, did not affect the sensitivity of yeast cells to rapamycin. However, the deletion of the CaHOG1 gene in Candida albicans can lead to cell sensitivity to rapamycin. These results suggest that Candida albicans HOG pathway is different from Saccharomyces cerevisiae HOG pathway in the regulation of cell function, which is involved in the regulation of cell sensitivity to rapamycin.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2009
【分類號】：R379

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