Turbidostat culture of Saccharomyces cerevisiae W303-1A under selective pressure elicited by ethanol selects for mutations in SSD1 and UTH1.
Turbidostat culture of Saccharomyces cerevisiae W303-1A under selective pressure elicited by ethanol selects for mutations in SSD1 and UTH1.
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DOI：
10.1111/j.1567-1364.2012.00803.x
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发表时间：
2012-08
影响因子：
3.2
通讯作者：
Braun S
中科院分区：
文献类型：
--
作者：
Avrahami-Moyal L;Engelberg D;Wenger JW;Sherlock G;Braun S
我们通过分析酿酒酵母W303-1A在乙醇选择压力下选择的突变来研究乙醇耐受性的遗传原因。W303-1A在培养基中添加越来越多的乙醇，进行三轮浊度控制。筛选结束时，培养物的生长速率由0.029 h-1提高到0.32 h-1。与前代菌株不同，从该群体中分离的所有酵母细胞都能够在添加7%乙醇的培养基上形成菌落，6天内，我们定义的乙醇耐受性。在添加9%乙醇的固体培养基上，三个阶段选择的几个无性系在两天内都能形成密集的菌落。我们对6个克隆的全基因组进行了测序，并确定了与乙醇耐受性有关的突变。另外13个克隆进行了类似突变的检测。在19个耐受性克隆中，有15个克隆ssd1-d的停止密码子被氨基酸编码密码子取代。另外三个克隆包含两个UTH1突变中的一个，一个克隆不包含SSD1或UTH1突变。我们发现，SSD1和UTH1的突变增加了细胞壁对酶解酶的耐受性，并得出结论，细胞壁的稳定性是增加对乙醇耐受性的主要因素。
We investigated the genetic causes of ethanol tolerance by characterizing mutations selected in Saccharomyces cerevisiae W303-1A under the selective pressure of ethanol. W303-1A was subjected to three rounds of turbidostat, in medium supplemented with increasing amounts of ethanol. By the end of selection, the growth rate of the culture has increased from 0.029 h-1 to 0.32 h-1. Unlike the progenitor strain, all yeast cells isolated from this population were able to form colonies on medium supplemented with 7% ethanol within six days, our definition of ethanol tolerance. Several clones selected from all three stages of selection were able to form dense colonies within two days on solid medium supplemented with 9% ethanol. We sequenced the whole genomes of 6 clones and identified mutations responsible for ethanol tolerance. Thirteen additional clones were tested for the presence of similar mutations. In 15 out of 19 tolerant clones the stop-codon in ssd1-d was replaced with an aminoacid-encoding codon. Three other clones contained one of two mutations in UTH1, and one clone did not contain mutations in either SSD1 or UTH1. We showed that the mutations in SSD1 and UTH1 increased tolerance of the cell wall to zymolyase and conclude that stability of the cell wall is a major factor in increased tolerance to ethanol.
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影响因子：
4.5
作者：
BENNETT, WN;BORAAS, ME
通讯作者：
BORAAS, ME
DOI：
10.1093/dnares/dsr029
发表时间：
2011-12
期刊：
DNA research : an international journal for rapid publication of reports on genes and genomes
影响因子：
--
作者：
Akao T;Yashiro I;Hosoyama A;Kitagaki H;Horikawa H;Watanabe D;Akada R;Ando Y;Harashima S;Inoue T;Inoue Y;Kajiwara S;Kitamoto K;Kitamoto N;Kobayashi O;Kuhara S;Masubuchi T;Mizoguchi H;Nakao Y;Nakazato A;Namise M;Oba T;Ogata T;Ohta A;Sato M;Shibasaki S;Takatsume Y;Tanimoto S;Tsuboi H;Nishimura A;Yoda K;Ishikawa T;Iwashita K;Fujita N;Shimoi H
通讯作者：
Shimoi H
影响因子：
3.6
作者：
Camougrand, N;Grelaud-Coq, A;Manon, S
通讯作者：
Manon, S
影响因子：
3.9
作者：
Camougrand, NM;Mouassite, M;Guérin, MG
通讯作者：
Guérin, MG
DOI：
10.1007/bf00500738
发表时间：
1982-01-01
期刊：
EUROPEAN JOURNAL OF APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
影响因子：
--
作者：
BROWN, SW;OLIVER, SG
通讯作者：
OLIVER, SG