Sampson J R, DiRenzo A B, Behlen L S, Uhlenbeck O C
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Biochemistry. 1990 Mar 13;29(10):2523-32. doi: 10.1021/bi00462a014.
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.
利用T7 RNA聚合酶进行体外转录，在参与酵母苯丙氨酸tRNA的9个三级碱基对或三联体的21个核苷酸中制备了32种不同的突变。这些突变的设计目的要么是破坏三级相互作用，要么是通过移植其他tRNA中存在的三级相互作用来改变序列而不破坏结构。用纯化的酵母苯丙氨酰合成酶进行的稳态氨酰化动力学研究表明，只要维持三级相互作用，反应速率几乎没有变化。这表明三级核苷酸仅有助于苯丙氨酸tRNA的折叠，而不直接参与与合成酶的序列特异性相互作用。
