A Saccharomyces cerevisiae genome-wide mutant screen for altered sensitivity to K1 killer toxin.
A Saccharomyces cerevisiae genome-wide mutant screen for altered sensitivity to K1 killer toxin.
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发表时间：
2003-03
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影响因子：
3.3
通讯作者：
N. Pagé;Manon Gérard-Vincent;Patrice Ménard;M. Beaulieu;M. Azuma;G. Dijkgraaf;Huijuan Li;José Marcoux;Thuy N Nguyen;Tim Dowse;A. Sdicu;H. Bussey
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作者：
N. Pagé;Manon Gérard-Vincent;Patrice Ménard;M. Beaulieu;M. Azuma;G. Dijkgraaf;Huijuan Li;José Marcoux;Thuy N Nguyen;Tim Dowse;A. Sdicu;H. Bussey
使用一组单独删除了 5718 个酵母基因的酿酒酵母突变体，我们筛选了对抗真菌蛋白 K1 杀伤毒素的敏感性改变，该毒素与细胞壁 β-葡聚糖受体结合，随后在质膜中形成致死孔。 268 个基因（包括 42 个功能未知的基因）的突变具有通常轻微的表型，与野生型相比，其中 186 个显示出耐药性，82 个显示出超敏反应。之前已知这些基因中只有 15 个在突变时会导致毒素表型。分析了 144 个基因突变体的碱溶性 β-葡聚糖水平； 63显示了改变。此外，还筛选了 118 个毒素敏感性改变的基因突变体，以检测 SDS、潮霉素 B 和 calcoflor White 敏感性，作为细胞表面缺陷的指标； 88 显示出一些额外的缺陷。突变体具有明显的非随机功能分布。许多基因影响细胞活动的特定区域，包括细胞壁葡聚糖和甘露糖蛋白合成、分泌途径运输、脂质和甾醇生物合成以及细胞表面信号转导，并为这些过程及其整合提供新的见解。
Using the set of Saccharomyces cerevisiae mutants individually deleted for 5718 yeast genes, we screened for altered sensitivity to the antifungal protein, K1 killer toxin, that binds to a cell wall beta-glucan receptor and subsequently forms lethal pores in the plasma membrane. Mutations in 268 genes, including 42 in genes of unknown function, had a phenotype, often mild, with 186 showing resistance and 82 hypersensitivity compared to wild type. Only 15 of these genes were previously known to cause a toxin phenotype when mutated. Mutants for 144 genes were analyzed for alkali-soluble beta-glucan levels; 63 showed alterations. Further, mutants for 118 genes with altered toxin sensitivity were screened for SDS, hygromycin B, and calcofluor white sensitivity as indicators of cell surface defects; 88 showed some additional defect. There is a markedly nonrandom functional distribution of the mutants. Many genes affect specific areas of cellular activity, including cell wall glucan and mannoprotein synthesis, secretory pathway trafficking, lipid and sterol biosynthesis, and cell surface signal transduction, and offer new insights into these processes and their integration.