Using canavanine resistance to measure mutation rates in Schizosaccharomyces pombe.
Using canavanine resistance to measure mutation rates in Schizosaccharomyces pombe.
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DOI：
10.1371/journal.pone.0271016
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
Kearsey, Stephen E.
中科院分区：
文献类型：
--
作者：
Pai, Chen-Chun;Heitzer, Ellen C.;Bertrand, Sibyl E.;Toumazou, Sophia;Humphrey, Timothy C.;Kearsey, Stephen E.
我们构建了一个S面板。pombe菌株表达与癌症相关的DNA聚合酶ε变体，特别是POLES 297 F，POLEV 411 L，POLEL 424 V，POLES 459 F，并使用这些来比较通过刀豆氨酸抗性与其他选择性方法确定的突变率。刀豆氨酸耐药突变率与ade-485突变逆转为腺嘌呤原养型的突变率大致相似，但低于5-氟乳清酸（FOA）耐药率（ura 4+或ura 5+基因失活）。几个基因的失活与S.粟酒裂殖酵母，但令人惊讶的全基因组测序显示，8/8自发刀豆氨酸抗性突变体在any 1/arn 1基因中具有R175 C突变。该基因编码一种α-抑制蛋白样蛋白，参与介导靶蛋白的Pub 1泛素化，该单一突变对刀豆氨酸的表型抗性与原始“can 1 -1”菌株所显示的相似，该菌株也具有any 1 R175 C突变。一些自发突变体在精氨酸转运蛋白中有额外的突变，这表明这可能会略微增加对刀豆氨酸的耐药性。any 1 R175 C菌株表现出Cat 1精氨酸转运蛋白的内化，解释了刀豆氨酸耐药表型。
We constructed a panel of S. pombe strains expressing DNA polymerase ε variants associated with cancer, specifically POLES297F, POLEV411L, POLEL424V, POLES459F, and used these to compare mutation rates determined by canavanine resistance with other selective methods. Canavanine-resistance mutation rates are broadly similar to those seen with reversion of the ade-485 mutation to adenine prototrophy, but lower than 5-fluoroorotic acid (FOA)-resistance rates (inactivation of ura4+ or ura5+ genes). Inactivation of several genes has been associated with canavanine resistance in S. pombe but surprisingly whole genome sequencing showed that 8/8 spontaneous canavanine-resistant mutants have an R175C mutation in the any1/arn1 gene. This gene encodes an α-arrestin-like protein involved in mediating Pub1 ubiquitylation of target proteins, and the phenotypic resistance to canavanine by this single mutation is similar to that shown by the original “can1-1” strain, which also has the any1R175C mutation. Some of the spontaneous mutants have additional mutations in arginine transporters, suggesting that this may marginally increase resistance to canavanine. The any1R175C strain showed internalisation of the Cat1 arginine transporter as previously reported, explaining the canavanine-resistance phenotype.
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常见的与癌症相关的DNA聚合酶ε突变会导致异常强的突变器表型，表明富达缺陷与校对丧失不同。
影响因子：
11.2
作者：
Kane DP;Shcherbakova PV
通讯作者：
Shcherbakova PV
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。
影响因子：
3.7
作者：
通讯作者：
--
影响因子：
2.4
作者：
Nakashima A;Kamada S;Tamanoi F;Kikkawa U
通讯作者：
Kikkawa U
影响因子：
7.7
作者：
Jiang P;Ollodart AR;Sudhesh V;Herr AJ;Dunham MJ;Harris K
通讯作者：
Harris K
影响因子：
3.5
作者：
Church DN;Briggs SE;Palles C;Domingo E;Kearsey SJ;Grimes JM;Gorman M;Martin L;Howarth KM;Hodgson SV;NSECG Collaborators;Kaur K;Taylor J;Tomlinson IP
通讯作者：
Tomlinson IP