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体外构建酵母天冬氨酸转运核糖核酸变体：T 茎和 T 环中的核苷酸替换与添加

In vitro construction of yeast tRNAAsp variants: nucleotide substitutions and additions in T-stem and T-loop.

作者信息

Carbon P, Ebel J P

出版信息

Nucleic Acids Res. 1987 Mar 11;15(5):1933-50. doi: 10.1093/nar/15.5.1933.

DOI:10.1093/nar/15.5.1933
PMID:3550694
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC340609/
Abstract

A procedure for the construction of 3'-end labelled yeast tRNAAsp harboring substitutions or additions of any desired nucleotide in T-stem and T-loop (position 57 to 61) has been developed. This was done by in vitro enzymatic manipulations of the yeast tRNAAsp involving specific hydrolysis with RNases, phosphorylation and dephosphorylation with T4 polynucleotide kinase and ligation with T4 RNA ligase. Using this procedure we have replaced conserved or semi-conserved nucleotides located in position 57 to 61 of yeast tRNAAsp. We have also constructed different yeast tRNAAsp with eight bases instead of seven in T-loop. Further use of these tRNAAsp variants will be discussed with the help of the crystallographic three-dimensional structure.

摘要

已开发出一种构建3'端标记的酵母天冬氨酸tRNA的方法，该tRNA在T茎和T环（第57至61位）中含有任何所需核苷酸的取代或添加。这是通过对酵母天冬氨酸tRNA进行体外酶促操作来完成的，包括用核糖核酸酶进行特异性水解、用T4多核苷酸激酶进行磷酸化和去磷酸化以及用T4 RNA连接酶进行连接。使用该方法，我们替换了酵母天冬氨酸tRNA第57至61位的保守或半保守核苷酸。我们还构建了不同的酵母天冬氨酸tRNA，其T环中有八个碱基而非七个碱基。将借助晶体学三维结构讨论这些天冬氨酸tRNA变体的进一步应用。

相似文献

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In vitro construction of yeast tRNAAsp variants: nucleotide substitutions and additions in T-stem and T-loop.体外构建酵母天冬氨酸转运核糖核酸变体：T 茎和 T 环中的核苷酸替换与添加
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2
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引用本文的文献

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Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.体外转录的未修饰酵母苯丙氨酸转移RNA的生化与物理特性
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Recognition of bases in Escherichia coli tRNA(Gln) by glutaminyl-tRNA synthetase: a complete identity set.

本文引用的文献

1
The effect of acceptor oligoribonucleotide sequence on the T4 RNA ligase reaction.受体寡核糖核苷酸序列对T4 RNA连接酶反应的影响。
Eur J Biochem. 1982 Jul;125(3):639-43. doi: 10.1111/j.1432-1033.1982.tb06730.x.
2
Reversal of T4 RNA ligase.T4 RNA连接酶的逆转
Biochemistry. 1982 Apr 13;21(8):1858-64. doi: 10.1021/bi00537a024.
3
Tertiary structure of tRNAs in solution monitored by phosphodiester modification with ethylnitrosourea.通过用乙基亚硝基脲进行磷酸二酯修饰监测溶液中tRNA的三级结构。
EMBO J. 1992 Nov;11(11):4159-65. doi: 10.1002/j.1460-2075.1992.tb05509.x.
Eur J Biochem. 1981 Sep;119(1):51-9. doi: 10.1111/j.1432-1033.1981.tb05575.x.
4
Conserved 5S rRNA complement to tRNA is not required for protein synthesis.蛋白质合成不需要与tRNA互补的保守5S rRNA。
Proc Natl Acad Sci U S A. 1982 Jan;79(1):36-40. doi: 10.1073/pnas.79.1.36.
5
Specific interaction of anticodon loop residues with yeast phenylalanyl-tRNA synthetase.反密码子环残基与酵母苯丙氨酰 - tRNA合成酶的特异性相互作用。
Biochemistry. 1982 Aug 17;21(17):3921-6. doi: 10.1021/bi00260a003.
6
Compilation of tRNA sequences.转运RNA序列的汇编。
Nucleic Acids Res. 1984;12 Suppl(Suppl):r1-57.
7
Enzymatic conversion of adenosine to inosine in the wobble position of yeast tRNAAsp: the dependence on the anticodon sequence.酵母天冬氨酸转运核糖核酸摆动位置上腺苷向次黄苷的酶促转化：对抗密码子序列的依赖性
Nucleic Acids Res. 1984 Mar 26;12(6):2705-15. doi: 10.1093/nar/12.6.2705.
8
Anticodon loop size and sequence requirements for recognition of formylmethionine tRNA by methionyl-tRNA synthetase.甲硫氨酰 - tRNA合成酶识别甲酰甲硫氨酸tRNA的反密码子环大小及序列要求。
Proc Natl Acad Sci U S A. 1983 Nov;80(22):6755-9. doi: 10.1073/pnas.80.22.6755.
9
Site-directed in vitro replacement of nucleosides in the anticodon loop of tRNA: application to the study of structural requirements for queuine insertase activity.tRNA反密码子环中核苷的定点体外置换：在对喹啉插入酶活性的结构要求研究中的应用
EMBO J. 1983;2(7):1093-7. doi: 10.1002/j.1460-2075.1983.tb01551.x.
10
Correlation between chemical modification and surface accessibility in yeast phenylalanine transfer RNA.酵母苯丙氨酸转运RNA中化学修饰与表面可及性之间的相关性
Biopolymers. 1983 Apr;22(4):1145-66. doi: 10.1002/bip.360220410.