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首页> 外文期刊>Nucleic acids research >Post-transcriptlonal modification of the wobble nucletide in anticodon-substituted yeast tRNAIIArg after microinjection into xenopus laevis oocytes
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Post-transcriptlonal modification of the wobble nucletide in anticodon-substituted yeast tRNAIIArg after microinjection into xenopus laevis oocytes

机译：在抗辐条取代的酵母Trnaiiarg中的发泡核苷酸后转录核细胞分解成Xenopus Laevis卵母细胞

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An enzymatic procedure for the replacement of the ICG anticodon of yeast tRNAλJg by NCG tnnucleotide (N=A, C, G or U) is described. Partial digestion with Si-nuclease and Ti-RNAase provides fragments which, when annealed together, form an “anticodon-deprived” yeast tRNA4?g. A novel anticodon, phosphorylated with (λ2P) label on its 51 terminal residue, is then inserted using T4-RNA ligase. Such “anticodon-substituted” yeast tRNAj* are micromjected into the cytoplasm of Xenopus laevis oocytes and shown to be able to interact with the anticodon maturation enzymes under in vivo conditions. Our results indicate that when adenosine occurs in the wobble position (A34***) in yeast tRNAλJg it is efficiently modified into mosine (I34) while uridine (U34) is transformed into two uridine derivatives, one of which is probably mcm5u. In contrast, when a cytosme (C34) or guanosine (G34) occurs, they are not modified. These results are at variance with those obtained previously under similar conditions with anticodon derivatives of yeast tRNAAsp harbouring A, C, G or U as the first anticodon nucleotide. In this case, guanosine and uridine were modified while adenosme and cytosme were not.
机译：描述了用于通过NCG叔核苷酸（n = A，C，G或U）替换酵母TRNAλjg的ICG抗oryon的酶促方法。用Si-核酸酶和Ti-RNA酶部分消化提供片段，当在一起时，将“抗助迁移的”酵母TrNA4〜G.然后使用T4-RNA连接酶在其5 1℃末端残留物上用（λ 2℃）标记磷酸化，用（λ 2 p）标记。这样的“抗助置的”酵母TrNAJ *是微微喷射到Xenopus Laevis卵母细胞的细胞质中，并显示能够与体内条件下的抗癌酶成熟酶相互作用。我们的结果表明，当腺苷发生在酵母TRNAλjg中发生腺苷（A34 ***）时，它被有效地修饰到莫宁（I34）中，而尿苷（U34）转化为两个尿素衍生物，其中可能是MCM5U。相反，当发生细胞菌（C34）或鸟苷（G34）时，它们不会被修改。这些结果与先前在类似条件下获得的结果的差异，含有酵母TrnaASP的抗oconon衍生物，其含有A，C，G或U作为第一抗病核苷酸。在这种情况下，腺苷和尿苷被修饰，而Adenosme和Cytosme则没有。

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