Heterologous expression and functional analysis of the F-box protein Ucc1 from other yeast species in Saccharomyces cerevisiae
Heterologous expression and functional analysis of the F-box protein Ucc1 from other yeast species in Saccharomyces cerevisiae
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DOI：
10.1016/j.jbiosc.2019.06.003
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发表时间：
2019-12-01
影响因子：
2.8
通讯作者：
Moriyama, Akihiko
中科院分区：
文献类型：
--
作者：
Nakatsukasa, Kunio;Kawarasaki, Tomoyuki;Moriyama, Akihiko
泛素-蛋白酶体系统在代谢调节中起着重要的作用。在以前的研究中，我们报告说，在酿酒酵母中，当葡萄糖是可用的，SCFUcc 1泛素连接酶复合物的目标柠檬酸合酶2（Cit 2）的蛋白酶体降解，从而抑制乙醛酸循环，一个合成代谢途径，取代TCA循环与琥珀酸的激活的代谢产物生成。然而，Ucc 1在其他酵母物种中的作用仍不清楚。在这里，我们克隆了直向同源的F-box蛋白Ucc 1从Zygosaccharomyces bailii，一个积极的食品腐败微生物，是最耐乙酸的酵母菌种，和光滑念珠菌，一种新兴的真菌病原体。这些同源基因在S. cerevisiae，并对其活性进行了遗传和生化测试。结果表明，Z. bailii Ucc 1拯救了ucc 1 Delta，表型，表明在Z.贝利。与此相反，C. glabrata Ucc 1不与ucc 1 Delta表型互补或对Ucc 1表现出显性负效应。这些结果表明，在广泛的酵母物种的乙醛酸循环的调控机制分析的重要性。(C)2019年，日本生物技术学会。All rights reserved.
The ubiquitin-proteasome system plays an important role in metabolic regulation. In a previous study, we reported that, in Saccharomyces cerevisiae, when glucose is available, the SCFUcc1 ubiquitin ligase complex targets citrate synthase 2 (Cit2) for proteasomal degradation, thereby suppressing the glyoxylate cycle, an anabolic pathway that replenishes the TCA cycle with succinate for the activation of gluconeogenesis. However, the roles of Ucc1 in other yeast species remain unclear. Here, we cloned orthologs of the F-box protein Ucc1 from Zygosaccharomyces bailii, an aggressive food spoilage microorganism that is the most acetic acid-tolerant yeast species, and Candida glabrata, an emerging fungal pathogen. These orthologs were expressed in S. cerevisiae, and their activities were tested genetically and biochemically. The results showed that Z. bailii Ucc1 rescued the ucc1 Delta, phenotype, suggesting the existence of a similar mechanism regulating the glyoxylate cycle in Z. bailii. By contrast, C. glabrata Ucc1 did not complement the ucc1 Delta phenotype or exhibit a dominant negative effect on Ucc1. These results suggest the importance of analysing the regulatory mechanisms of glyoxylate cycle in a broad range of yeast species. (C) 2019, The Society for Biotechnology, Japan. All rights reserved.