Role of the tertiary nucleotides in the interaction of yeast phenylalanine tRNA with its cognate synthetase.
Role of the tertiary nucleotides in the interaction of yeast phenylalanine tRNA with its cognate synthetase.
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DOI：
10.1021/bi00462a014
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发表时间：
1990-03
期刊：
影响因子：
2.9
通讯作者：
J. Sampson;A. Direnzo;L. Behlen;O. Uhlenbeck
中科院分区：
文献类型：
--
作者：
J. Sampson;A. Direnzo;L. Behlen;O. Uhlenbeck
通过T7 RNA聚合酶的体外转录用于制备参与酵母tRNAPhe的9个三级碱基对或三联体的21个核苷酸中的32个不同突变。突变被设计为破坏三级相互作用或通过移植存在于其他tRNA中的三级相互作用来改变序列而不破坏结构。稳态氨酰化动力学与纯化的酵母苯丙氨酰合成酶的反应速率变化不大，只要三级相互作用保持。这表明三级核苷酸仅有助于tRNAPhe的折叠，并且不直接参与与合成酶的序列特异性相互作用。
In vitro transcription by T7 RNA polymerase was used to prepare 32 different mutations in the 21 nucleotides that participate in the 9 tertiary base pairs or triples of yeast tRNAPhe. The mutations were designed either to disrupt the tertiary interaction or to change the sequence without disrupting the structure by transplanting tertiary interactions present in other tRNAs. Steady-state aminoacylation kinetics with purified yeast phenylalanyl synthetase revealed little change in reaction rate as long as a tertiary interaction was maintained. This suggests that the tertiary nucleotides only contribute to the folding of tRNAPhe and do not participate directly in sequence-specific interaction with the synthetase.