rDNA Copy Number Variants Are Frequent Passenger Mutations in Saccharomyces cerevisiae Deletion Collections and de Novo Transformants.
rDNA Copy Number Variants Are Frequent Passenger Mutations in Saccharomyces cerevisiae Deletion Collections and de Novo Transformants.
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DOI：
10.1534/g3.116.030296
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发表时间：
2016-09-08
期刊：
影响因子：
--
通讯作者：
Raghuraman MK
中科院分区：
文献类型：
--
作者：
Kwan EX;Wang XS;Amemiya HM;Brewer BJ;Raghuraman MK
已知酿酒酵母核糖体DNA（rDNA）基因座相对于基因组的其余部分表现出更大的不稳定性。然而，野生型细胞优先保持稳定的rDNA拷贝数，这表明该基因座的大小的潜在遗传控制。我们进行了一个子集的酵母敲除（YKO）单基因缺失收集的筛选，以确定该基因座的遗传调节因子，并确定rDNA拷贝数是否与酵母复制寿命相关。虽然我们没有发现复制寿命和rDNA大小之间的相关性，我们确定了64个候选菌株具有显着的rDNA拷贝数差异。然而，在验证候选rDNA变体的过程中，我们观察到我们的从头基因缺失菌株的独立分离株在rDNA拷贝数上有自发但显著的变化。此外，我们不能概括来自YKO酵母缺失集合的rDNA表型。相反，我们发现标准乙酸锂转化方案是rDNA拷贝数变异的重要来源，乙酸锂暴露是转化后引起可变rDNA拷贝数事件的处理。随着rDNA拷贝数变异的影响越来越多地被报道，我们发现rDNA受到醋酸锂暴露的影响，这表明rDNA拷贝数变异可能是S.啤酒。
The Saccharomyces cerevisiae ribosomal DNA (rDNA) locus is known to exhibit greater instability relative to the rest of the genome. However, wild-type cells preferentially maintain a stable number of rDNA copies, suggesting underlying genetic control of the size of this locus. We performed a screen of a subset of the Yeast Knock-Out (YKO) single gene deletion collection to identify genetic regulators of this locus and to determine if rDNA copy number correlates with yeast replicative lifespan. While we found no correlation between replicative lifespan and rDNA size, we identified 64 candidate strains with significant rDNA copy number differences. However, in the process of validating candidate rDNA variants, we observed that independent isolates of our de novo gene deletion strains had unsolicited but significant changes in rDNA copy number. Moreover, we were not able to recapitulate rDNA phenotypes from the YKO yeast deletion collection. Instead, we found that the standard lithium acetate transformation protocol is a significant source of rDNA copy number variation, with lithium acetate exposure being the treatment causing variable rDNA copy number events after transformation. As the effects of variable rDNA copy number are being increasingly reported, our finding that rDNA is affected by lithium acetate exposure suggested that rDNA copy number variants may be influential passenger mutations in standard strain construction in S. cerevisiae.
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影响因子：
4.5
作者：
Ide S;Saka K;Kobayashi T
通讯作者：
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DOI：
10.1073/pnas.75.4.1929
发表时间：
1978-01-01
影响因子：
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作者：
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影响因子：
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作者：
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