Unlinked noncomplementation: isolation of new conditional-lethal mutations in each of the tubulin genes of Saccharomyces cerevisiae.
Unlinked noncomplementation: isolation of new conditional-lethal mutations in each of the tubulin genes of Saccharomyces cerevisiae.
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DOI：
10.1093/genetics/119.2.249
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发表时间：
1988-06
期刊：
影响因子：
3.3
通讯作者：
T. Stearns;D. Botstein
中科院分区：
文献类型：
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作者：
T. Stearns;D. Botstein
通过筛选不能互补单个β-微管蛋白编码基因（TUB 2）中突变的非连锁突变，寻找编码与β-微管蛋白相互作用的蛋白质的酿酒酵母基因中的突变。在检查的前三个非互补突变中，两个与TUB 2相关，而一个与TUB 2无关。该非连锁突变被证明是主要α-微管蛋白编码基因（TUB 1）的条件致死等位基因，并代表该基因中的第一个此类突变。tub 1 -1突变本身导致冷敏感细胞周期停滞，并赋予抗微管药物苯菌灵超敏性。这些表型发生在一个野生型拷贝的次要α-微管蛋白编码基因，TUB 3的存在下，tub 1 -1和tub 3无效突变的组合是不可活的单倍体。通过进一步应用这种方法，TUB 2和TUB 3中的新突变被分离为tub 1 -1的非连锁非互补物。tub 1和tub 2突变之间的非互补是基因特异性和等位基因特异性的，表明表型是由于在蛋白质水平上的相互作用。我们的结论是，分离不连锁的非互补突变可能是一个普遍有用的方法，分离相互作用的基因产物中的突变。
Mutations in genes of Saccharomyces cerevisiae that code for proteins that interact with beta-tubulin were sought by screening for unlinked mutations that fail to complement mutations in the single beta-tubulin-encoding gene (TUB2). Among the first three noncomplementing mutations examined, two are linked to TUB2 while one is unlinked. The unlinked mutation was shown to be a conditional-lethal allele of the major alpha-tubulin-encoding gene (TUB1) and represents the first such mutation in that gene. The tub1-1 mutation itself causes a cold-sensitive cell-cycle arrest, and confers supersensitivity to the antimicrotubule drug benomyl. These phenotypes occur in the presence of a wild-type copy of the minor alpha-tubulin-encoding gene, TUB3; the combination of tub1-1 and a tub3 null mutation is inviable in haploids. Through further application of this method, new mutations in TUB2 and TUB3 were isolated as unlinked noncomplementers of tub1-1. The noncomplementation between tub1 and tub2 mutations is gene specific and allele specific, suggesting that the phenotype is due to an interaction at the protein level. We conclude that isolation of unlinked noncomplementing mutations is likely to be a generally useful method for isolating mutations in interacting gene products.