ALPHA-DNA-VI - COMPARATIVE-STUDY OF ALPHA-ANOMERIC AND BETA-ANOMERIC OLIGODEOXYRIBONUCLEOTIDES IN HYBRIDIZATION TO MESSENGER-RNA AND IN CELL FREE TRANSLATION INHIBITION
ALPHA-DNA-VI - COMPARATIVE-STUDY OF ALPHA-ANOMERIC AND BETA-ANOMERIC OLIGODEOXYRIBONUCLEOTIDES IN HYBRIDIZATION TO MESSENGER-RNA AND IN CELL FREE TRANSLATION INHIBITION
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DOI：
10.1093/nar/15.24.10419
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发表时间：
1987-12-23
影响因子：
14.9
通讯作者：
PAOLETTI, C
中科院分区：
文献类型：
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作者：
GAGNOR, C;BERTRAND, JR;PAOLETTI, C
比较了α和α-异头d（G2 T12 G2）寡脱氧核糖核苷酸与rA 12的杂交：α-寡脱氧核糖核苷酸/RNA杂交体的观察到的解链温度为27°C，α-寡脱氧核糖核苷酸/RNA为53°C。首次合成了与天然mRNA互补的4个碱基的α-寡核苷酸，并在其5′端用[32 P]标记，用作北方印迹实验的探针。尽管它们对靶RNA具有较高的亲和力，但它们不能阻断兔网织红细胞裂解物中天然或合成mRNA的翻译。我们研究了RNA酶H在rA 12：α-或α-d（G2 T12 G2）杂交模型或mRNA：α-或α-寡核苷酸杂交模型上的活性。当使用α-寡核苷酸而不是β-寡核苷酸时，特异性杂交保护RNA拉伸。因此，我们的结果表明RNase H不能降解α-寡脱氧核糖核苷酸：RNA双链体中的RNA。
α and ß-anomeric d(G2T12G2) oligodeoxyribonucleotides were compared for their hybridization to rA12: the observed melting temperatures are 27°C for ß-oligodeoxyribonucleotide/RNA hybrid and 53°C for α-oligodeoxyribonucleotide/RNA. α-oligonucleotides with the four bases, complementary to natural mRNAs, were synthesized for the first time, labeled at their 5′-end with [32P] and used as probes in Northern blot experiments. In spite of these higher affinities for their target RNA's, they were unable to block translation of natural or synthetic mRNA's in rabbit reticulocyte lysate. We have studied the RNase H activity on model rA12:α- or ß-d(G2T12G2) hybrids or on mRNA:α- or ß-oligonucleotides hybrids. Specific hybridization protects RNA strech when using α-oligonucleotides but not ß-oligonucleotides. Thus, our results show the inability of RNase H to degrade RNA in α-oligodeoxyribonucleotides:RNA duplexes.