Raftery L A, Yarus M
EMBO J. 1987 May;6(5):1499-506. doi: 10.1002/j.1460-2075.1987.tb02392.x.
Using site-specific mutagenesis, we constructed five more efficient variants of tRNA(Glu)-Suoc, an extremely inefficient ochre suppressor. Each variant has an extended anticodon, or region of the anticodon arm, which is more similar to that found in normal tRNAs which translate codons Uxx. Suppressor efficiency invariably increases with similarity of the extended anticodon to that of a normal Uxx-translating tRNA. Altered nucleotides in both helix and loop strongly affect efficiency, with no position dependence and no significant interaction between substitutions. The variant with all substitutions is 230-fold more efficient (in one context) than the parental tRNA(Glu)-Suoc. Two other unexpected variants seem to be 'context mutants', having altered response to message context.
通过位点特异性诱变,我们构建了5个效率更高的tRNA(Glu)-Suoc变体,tRNA(Glu)-Suoc是一种效率极低的赭石抑制子。每个变体都有一个延长的反密码子,或反密码子臂区域,它与正常翻译密码子Uxx的tRNA中的反密码子臂区域更为相似。抑制子效率总是随着延长的反密码子与正常翻译Uxx的tRNA的反密码子之间的相似性增加而提高。螺旋和环中改变的核苷酸强烈影响效率,不存在位置依赖性,替代之间也没有显著的相互作用。在一种情况下,具有所有替代的变体比亲本tRNA(Glu)-Suoc的效率高230倍。另外两个意外的变体似乎是“上下文突变体”,对信息上下文的反应有所改变。