酿酒酵母菌和白念珠菌中RCK2和HOG1蛋白激酶在高渗胁迫，氧化胁迫和细胞壁完整性方面的功能研究
发布时间：2018-08-03 20:08
【摘要】： 酿酒酵母ScRck2p是一种MAP Knase激活的蛋白激酶,被Hog1p磷酸化而激活,响应于细胞对外界高渗透压胁迫和氧化胁迫。在本工作中我们发现酿酒酵母ScRCK2基因的缺失能够导致细胞对TOR抑制剂——雷帕霉素敏感,但ScRCK2在细胞对雷帕霉素敏感过程中的作用并不依赖于其激酶活性。在人体内最常见的机会致病菌-白念珠菌中,我们鉴定了ScRCK2的同源基因CaRCK2,并发现CaRCK2基因的缺失同样导致白念珠菌细胞对雷帕霉素敏感,CaRck2p的这一作用同样不依赖于其激酶活性。此外,我们发现白念珠菌CaHOG1的缺失也导致细胞对雷帕霉素的敏感。这些结果表明在酿酒酵母菌和白念珠菌中,作用于HOG途径下游的RCK2可能调控TOR信号途径的功能。 同时,我们发现白念珠菌CaRCK2基因的缺失能够导致细胞对高渗胁迫、氧化胁迫和细胞壁完整性胁迫试剂敏感。比较而言,酿酒酵母ScRCK2基因的缺失只引起细胞对氧化胁迫敏感,而不影响细胞对高渗胁迫和细胞壁完整性胁迫试剂的敏感。但是,酿酒酵母ScRCK2基因却能够弥补白念珠菌CaRCK2基因缺失株对高渗胁迫、氧化胁迫和细胞壁完整性胁迫试剂的敏感性表型。这些结果说明RCK2在细胞内发挥多种功能,但在白念珠菌和酿酒酵母菌中的功能存在分歧。 我们还发现,ScRCK2上游激酶基因ScHOG1和ScPBS2的缺失,以及受ScHOG1调控的转录因子基因ScHOT1,ScMSN1,ScMSN2,ScMSN4或ScRLM1的缺失,不影响酵母细胞对雷帕霉素的敏感性。但是,白念珠菌CaHOG1基因的缺失能够导致细胞对雷帕霉素敏感。这些结果表明白念珠菌HOG途径与酿酒酵母HOG途径在细胞功能调控方面存在分歧,它参与细胞对雷帕霉素敏感性的调控。
[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.
【学位授予单位】：天津大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R379
[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.
【学位授予单位】：天津大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R379
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