Fluorine-19 nuclear magnetic resonance study of codon-anticodon interaction in 5-fluorouracil-substituted E. coli transfer RNAs.
Fluorine-19 nuclear magnetic resonance study of codon-anticodon interaction in 5-fluorouracil-substituted E. coli transfer RNAs.
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5-氟尿嘧啶取代的大肠杆菌转移 RNA 中密码子-反密码子相互作用的氟 19 核磁共振研究。
DOI：
10.1093/nar/14.11.4659
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发表时间：
1986
影响因子：
14.9
通讯作者：
Horowitz,J
中科院分区：
文献类型：
--
作者：
Gollnick,P;Hardin,CC;Horowitz,J
用~(19)F核磁共振波谱研究了全活性5-氟尿嘧啶取代的E.coll-tRNAYal(反密码子FAC)中的密码子相互作用。密码子GpUpA的结合导致19F核磁共振谱中心区域3.9ppm的19F共振前场位移，而不与反密码子互补的三核苷酸不起作用。同样的19F共振在19F标记的tRNA和E.coll tRNA2Tyr(反密码子qua)之间形成反密码子-反密码子二聚体时前移。这些结果允许将3.9ppm的峰指定给氟尿嘧啶取代的tRNA1Ya1反密码子中第34位的5-氟尿嘧啶。甲硫氨酸密码子ApUpG还导致含氟大肠杆菌tRNAmMet的19F核磁共振谱中央部分的峰发生序列特异性的前场移位。然而，ApUpG对19F标记的大肠杆菌tRNArMet的19F谱没有影响，表明启动子的反密码环和延长链的蛋氨酸tRNA之间可能存在构象差异。19F核磁共振实验没有检测到CpGpApA与5-氟尿嘧啶取代的tRNA1Yal的T环中互补的FpFpCpG(取代TPψpCpG)结合，这表明T环和D环之间的第三级相互作用没有被密码子-反密码子相互作用破坏。
Codonanticodon interaction was investigated in fully active 5-fluorouracil-substituted E. coll tRNAYal(anticodon FAC) by 19FNMR spectroscopy. Binding of the codon GpUpA results in the upfield shift of a19F resonance at 3.9'ppm in the central region of the19F NMR spectrum, whereas trinucleotides not complementary to the anticodon have no effect. The same19F resonance shifts upfield upon formation of an anticodon-anticodon dimer between the19F-labeled tRNA and E. coll tRNA2Tyr(anticodon QUA). These results permit assignment of the peak at 3.9 ppm to the 5-fluorouracil at position 34 in the anticodon of fluorouracil-substituted tRNA1Ya1. The methionine codon ApUpG also causes a sequence-specific upfield shift of a peak in the central part of the19F NMR spectrum of fluorinated E. coli tRNAmMet. However, ApUpG has no effect on the19F spectrum of19F-labeled E. coli tRNArMet, indicating possible conformational differences between the anti-codon loop of initiator and chain-elongating methionine tRNAs.19F NMR experiments detect no binding of CpGpApA to the complementary FpFpCpG (replaces TpψpCpG) in the T-loop of 5-fluorouracil-substituted tRNA1Yal, in the presence or absence of codon, suggesting that the tertiary interactions between the T- and D-loops are not disrupted by codon-anticodon interactions.