Characterization of canavanine-resistance of cat1 and vhc1 deletions and a dominant any1 mutation in fission yeast.
Characterization of canavanine-resistance of cat1 and vhc1 deletions and a dominant any1 mutation in fission yeast.
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CAT1和VHC1缺失的加拿大抗性的表征以及裂变酵母中的主要Any1突变。
DOI：
10.1371/journal.pone.0269276
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
正选择和反选择标记已成功地整合为许多模式生物的许多遗传分析的一部分。本研究探讨了pombe Schizosaccharomyces对精氨酸类似物菜豆氨酸的抗性机制及其在遗传选择中的适用性。精氨酸渗透酶基因cat1和SPBC18H10.16/vhc1（以前错误地称为can1）的缺失都提供了很强的耐药性，而SPBC18H10.16/vhc1的单个缺失对菜豆氨酸耐药性没有影响。令人惊讶的是，广泛使用的can1-1等位基因并不编码有缺陷的精氨酸渗透酶，而是对应于any1-523C >t等位基因。这种等位基因具有很强的抗大麻碱能力，因为它不能在细胞膜上沉积碱性氨基酸转运体。Any1 - 523c >T导致Tor2激酶调控的Any1翻译后修饰减少。我们还证明了any1- 523c>t是显性等位基因。我们的研究结果揭示了裂变酵母对大麻碱抗性的机制，并为在遗传分析中使用cat1， vhc1和any1突变等位基因开辟了机会。
Positive and counter-selectable markers have been successfully integrated as a part of numerous genetic assays in many model organisms. In this study, we investigate the mechanism of resistance to arginine analog canavanine and its applicability for genetic selection in Schizosaccharomyces pombe. Deletion of both the arginine permease gene cat1 and SPBC18H10.16/vhc1 (formerly mistakenly called can1) provides strong drug resistance, while the single SPBC18H10.16/vhc1 deletion does not have an impact on canavanine resistance. Surprisingly, the widely used can1-1 allele does not encode for a defective arginine permease but rather corresponds to the any1-523C>T allele. The strong canavanine-resistance conferred by this allele arises from an inability to deposit basic amino acid transporters on the cellular membrane. any1-523C>T leads to reduced post-translational modifications of Any1 regulated by the Tor2 kinase. We also demonstrate that any1-523C>T is a dominate allele. Our results uncover the mechanisms of canavanine-resistance in fission yeast and open the opportunity of using cat1, vhc1 and any1 mutant alleles in genetic assays.
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