Research Institute for Bioresources, Okayama University, 2-20-1, Chuo, Kurashiki, Okayama, 710-0046, Japan.
J Biosci Bioeng. 2010 May;109(5):453-8. doi: 10.1016/j.jbiosc.2009.10.015.
Rhodotorula glutinis IFO1125 acquired increased aluminum (Al) resistance from 50 muM to more than 5 mM by repetitive culturing with stepwise increases in Al concentration at pH 4.0. In our previous study we performed differential display to find that three genes (RgFET3, RgGET3, and RgCMK) encoding proteins homologous to Saccharomyces cerevisiae FET3p, GET3p, and CMK1p and CMK2p, respectively, were up-regulated in the Al-resistant cells. In this study we cloned these genes and found they were indeed up-regulated in Al-resistant strains. The cloned genes were introduced into S. cerevisiae and corresponding mutants to test their relevance to Al resistance. The introduction of RgFET3 and RgGET3 conferred Al resistance to the host, but that of RgCMK did not. Green fluorescent protein (GFP)-tagged RgFet3p was localized at the cell periphery in the host. GFP-tagged RgGet3p formed more punctate bodies in the host under Al stress than in the absence of Al. Different growth responses to Fe (III), Cu (II), Ca ions, and cyclosporin A in the wild type and resistant cells of R. glutinis suggested the involvement and possible links of the three genes in Al resistance.
粘红酵母IFO1125 在 pH4.0 下通过逐步增加铝浓度进行重复培养，从 50µM 增加到 5mM 以上，获得了增强的铝（Al）抗性。在我们之前的研究中，我们进行了差异显示，发现三个基因（RgFET3、RgGET3 和 RgCMK）编码的蛋白质与酿酒酵母 FET3p、GET3p 和 CMK1p 和 CMK2p 同源，在耐铝细胞中上调。在本研究中，我们克隆了这些基因，并发现它们在耐铝菌株中确实上调。将这些克隆的基因导入酿酒酵母及其相应的突变体，以测试它们与 Al 抗性的相关性。RgFET3 和 RgGET3 的引入赋予了宿主 Al 抗性，但 RgCMK 的引入没有。绿色荧光蛋白（GFP）标记的 RgFet3p 在宿主中定位于细胞边缘。在 Al 胁迫下，GFP 标记的 RgGet3p 在宿主中形成的点状体比在没有 Al 时更多。野生型和粘红酵母耐铝细胞对 Fe（III）、Cu（II）、Ca 离子和环孢菌素 A 的不同生长反应表明，这三个基因参与并可能与 Al 抗性有关。
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