Identification and dissection of a complex DNA repair sensitivity phenotype in Baker's yeast.
Identification and dissection of a complex DNA repair sensitivity phenotype in Baker's yeast.
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DOI：
10.1371/journal.pgen.1000123
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发表时间：
2008-07-11
期刊：
影响因子：
4.5
通讯作者：
Alani, Eric
中科院分区：
文献类型：
--
作者：
Demogines, Ann;Smith, Erin;Kruglyak, Leonid;Alani, Eric
复杂性状通常涉及多个基因变异的贡献。在这项研究中，我们利用高密度的基因分型分析的BY（S288 c）和RM菌株的酿酒酵母和123个衍生的孢子后代，以确定一个复杂的DNA修复敏感性表型的遗传位点。这是通过筛选杂交酵母后代对各种DNA损伤剂的敏感性来实现的。BY和RM菌株都对紫外线模拟剂4-硝基喹啉1-氧化物（4-NQO）有抗性;然而，BY×RM杂交的杂交后代对该药物表现出不同的敏感性。我们绘制了一个主要的数量性状位点（QTL），RAD 5，并确定了在这个位点内负责4-NQO敏感性的确切多态性。通过使用回交策略沿着阵列辅助批量分离分析，我们确定了另一个位点，MKT 1，和染色体VII上的QTL，也链接到杂交4-NQO敏感表型，但赋予更小的影响。这项工作提出了一个加性模型的敏感性4-NQO，并提供了一个战略，定位主要和次要QTL，赋予背景特异性表型。它还提供了了解遗传背景对遗传毒性剂敏感性的影响的工具。由于多个基因变异的贡献，复杂性状通常显示一系列表型。统计模型、遗传作图、DNA基因分型和测序的进展使面包酵母成为鉴定数量性状基因座（QTL）的优秀系统，QTL是与数量表型性状相关的基因组区域。我们专注于一个复杂的DNA损伤敏感性表型的酵母，其中亲本菌株不受影响，但产生后代的敏感表型。我们使用全基因组方法分离出两株面包酵母中基因变异引起的DNA修复缺陷，这两株面包酵母显示出约0.5%的序列差异。亲本菌株对紫外光模拟剂4-硝基喹啉1-氧化物（4-NQO）具有抗性;然而，大量孢子后代对该药物表现出不同的敏感性。通过连锁和批量分离分析，我们确定了一个主要的QTL，RAD 5，和两个次要的QTL连锁的敏感性4-NQO，我们提供的证据表明，敏感性是由于涉及几个QTL的加性效应。这些观察结果提供了一个强大的模型，在其中了解疾病的发病率的基础，以及如何表型变异可以在基因水平上定位。
Complex traits typically involve the contribution of multiple gene variants. In this study, we took advantage of a high-density genotyping analysis of the BY (S288c) and RM strains of Saccharomyces cerevisiae and of 123 derived spore progeny to identify the genetic loci that underlie a complex DNA repair sensitivity phenotype. This was accomplished by screening hybrid yeast progeny for sensitivity to a variety of DNA damaging agents. Both the BY and RM strains are resistant to the ultraviolet light–mimetic agent 4-nitroquinoline 1-oxide (4-NQO); however, hybrid progeny from a BY×RM cross displayed varying sensitivities to the drug. We mapped a major quantitative trait locus (QTL), RAD5, and identified the exact polymorphism within this locus responsible for 4-NQO sensitivity. By using a backcrossing strategy along with array-assisted bulk segregant analysis, we identified one other locus, MKT1, and a QTL on Chromosome VII that also link to the hybrid 4-NQO–sensitive phenotype but confer more minor effects. This work suggests an additive model for sensitivity to 4-NQO and provides a strategy for mapping both major and minor QTL that confer background-specific phenotypes. It also provides tools for understanding the effect of genetic background on sensitivity to genotoxic agents. Complex traits often display a range of phenotypes due to the contribution of multiple gene variants. Advances in statistical models, genetic mapping, and DNA genotyping and sequencing have made baker's yeast an excellent system to identify quantitative trait loci (QTL), regions of the genome linked to a quantitative phenotypic trait. We focused on a complex DNA damage sensitivity phenotype in yeast in which parental strains are unaffected but give rise to progeny with a sensitive phenotype. We used a whole-genome approach to isolate defects in DNA repair caused by gene variants in two strains of baker's yeast that display approximately 0.5% sequence divergence. The parental strains are resistant to the ultraviolet light–mimetic agent 4-nitroquinoline 1-oxide (4-NQO); however, a large number of spore progeny displayed varying sensitivities to the drug. Through linkage and bulk segregant analyses we identified one major QTL, RAD5, and two minor QTL linked to sensitivity to 4-NQO, and we provide evidence that sensitivity is due to additive effects involving several QTL. These observations provide a powerful model in which to understand the basis of disease penetrance and how phenotypic variation can be mapped at the gene level.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
DOI：
10.1073/pnas.0510998103
发表时间：
2006-02-28
影响因子：
11.1
作者：
Heck, JA;Argueso, JL;Alani, E
通讯作者：
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影响因子：
56.9
作者：
Gresham, D;Ruderfer, DM;Kruglyak, L
通讯作者：
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影响因子：
5.8
作者：
Broman, KW;Wu, H;Churchill, GA
通讯作者：
Churchill, GA
DOI：
10.1073/pnas.262669299
发表时间：
2002-12-24
影响因子：
11.1
作者：
Chang, M;Bellaoui, M;Brown, GW
通讯作者：
Brown, GW