Masselot M, Robichon-Szulmajster H
Genetics. 1972 Aug;71(4):535-50. doi: 10.1093/genetics/71.4.535.
Ethionine-resistant mutants, mapping at the locus eth2-the product of which is involved in pleiotropic regulation of methionine biosynthesis-have been isolated in a strain carrying five ochre nonsense mutations. Selection for nonsense suppressors in such a strain led to characterization of several allele-specific but gene non-specific suppressors which are active on the recessive heteroallele eth2-2 (resulting in partial recovery of sensitivity toward ethionine) as well as on the five other suppressible alleles. Two of these suppressors are unlinked to the eth2 gene and either dominant or semi-dominant. It is concluded that the mutation eth2-2 resulted in a nonsense codon. Enzyme studies indicate that this mutation results in a complete absence of an active product of gene eth2, in contrast with the effect of a former mutation eth2-1 which was interpreted as leading to a modified product of this gene (Cherest, Surdin-Kerjan and de Robichon-Szulmajster 1971). This conclusion is based on the absence of repressibility of methionine group I enzymes and the observation that in a heteroallelic diploid, eth2-1 expression is not masked by eth2-2. The nonsense suppressors studied lead to at least partial recovery of repressibility of methionine group I enzymes. All these results support the idea that the product of gene ETH2 is an aporepressor protein.
在携带五个赭石型无义突变的菌株中,分离出了抗乙硫氨酸突变体,这些突变体定位于eth2位点,其产物参与甲硫氨酸生物合成的多效性调控。在这样的菌株中选择无义抑制子,导致鉴定出几种等位基因特异性但基因非特异性的抑制子,它们对隐性异等位基因eth2-2(导致对乙硫氨酸的敏感性部分恢复)以及其他五个可抑制等位基因都有活性。其中两个抑制子与eth2基因不连锁,要么是显性的,要么是半显性的。得出的结论是,eth2-2突变产生了一个无义密码子。酶学研究表明,与之前的eth2-1突变(被解释为导致该基因的修饰产物)的效应相反,这个突变导致eth2基因的活性产物完全缺失(Cherest、Surdin-Kerjan和de Robichon-Szulmajster,1971)。这个结论基于甲硫氨酸I组酶缺乏可阻遏性,以及在异等位基因二倍体中观察到eth2-1的表达不被eth2-2掩盖。所研究的无义抑制子导致甲硫氨酸I组酶的可阻遏性至少部分恢复。所有这些结果都支持基因ETH2的产物是一种无辅基阻遏蛋白这一观点。