Farofonova Vasilina, Karginov Azamat, Zvonarev Anton, Kulakovskaya Ekaterina, Agaphonov Michael, Kulakovskaya Tatiana
Federal Research Center "Pushchino Scientific Center for Biological Research", Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino, Russian Federation.
Bach Institute of Biochemistry, Federal Research Center "Fundamentals of Biotechnology" of the Russian Academy of Sciences, Moscow, Russian Federation.
Folia Microbiol (Praha). 2024 Dec 27. doi: 10.1007/s12223-024-01236-2.
Cells of the methylotrophic yeast Ogataea parapolymorpha have two genes encoding low-affinity phosphate transporters: PHO87, encoding the plasma membrane transporter, and PHO91, encoding a protein, which is homologous to the Saccharomyces cerevisiae vacuolar membrane transporter. Earlier, we reported that inactivation of PHO91 in O. parapolymorpha interferes with methanol utilization due to the lack of activity of methanol oxidase encoded by the MOX gene. In this work, we showed that this defect was completely suppressed by inactivating the PHO87 gene or introducing additional copies of the MOX gene into the cell. The PHO91 gene knockout decreased the level of long-chained polyphosphates only in methanol-grown cells, but not in glucose-grown cells. This effect remained even in the strain with extra copies of MOX, which rescues the ability of the mutant to grow on methanol. In contrast, the PHO87 gene knockout changed the levels of short-chained and long-chained polyphosphates in both methanol- and glucose-grown cells. Inactivation of PHO91 did not change vanadate resistance, while inactivation of PHO87 increased this resistance. Our data suggest that in O. parapolymorpha, Pho87 and Pho91 transporters have different roles in inorganic polyphosphate metabolism and adaptation to methanol consumption.
甲基营养型酵母嗜甲醇毕赤酵母(Ogataea parapolymorpha)细胞有两个编码低亲和力磷酸盐转运蛋白的基因:PHO87,编码质膜转运蛋白;PHO91,编码一种与酿酒酵母(Saccharomyces cerevisiae)液泡膜转运蛋白同源的蛋白质。此前,我们报道嗜甲醇毕赤酵母中PHO91的失活会干扰甲醇利用,原因是MOX基因编码的甲醇氧化酶缺乏活性。在这项工作中,我们表明通过失活PHO87基因或将MOX基因的额外拷贝引入细胞,这种缺陷能被完全抑制。PHO91基因敲除仅降低了甲醇培养细胞中长链多聚磷酸盐的水平,而在葡萄糖培养细胞中则没有。即使在具有MOX额外拷贝的菌株中这种效应仍然存在,该菌株挽救了突变体在甲醇上生长的能力。相反,PHO87基因敲除改变了甲醇和葡萄糖培养细胞中短链和长链多聚磷酸盐的水平。PHO91失活不会改变钒酸盐抗性,而PHO87失活会增加这种抗性。我们的数据表明,在嗜甲醇毕赤酵母中,Pho87和Pho91转运蛋白在无机多聚磷酸盐代谢和对甲醇消耗的适应中具有不同作用。