含O6-甲基鸟苷的寡核糖核苷酸的合成：锤头状核酶切割中保守鸟苷残基的作用
The synthesis of oligoribonucleotides containing O6-methylguanosine: the role of conserved guanosine residues in hammerhead ribozyme cleavage.
作者信息
Grasby J A, Jonathan P, Butler G, Gait M J
机构信息
MRC Laboratory of Molecular Biology, Cambridge, UK.
出版信息
Nucleic Acids Res. 1993 Sep 25;21(19):4444-50. doi: 10.1093/nar/21.19.4444.
Abstract
The synthesis is described of oligoribonucleotides containing the modified nucleoside O6-methylguanosine. Solid-phase oligoribonucleotide assembly was carried out by use of 2'-silyl-protected nucleoside phosphoramidites, a new O6-methylguanosine-containing synthon and a mild deprotection method. The O6-methylguanosine-modified oligonucleotides were used in the study of the role of conserved residues G5, G8 and G12 in hammerhead ribozyme cleavage. Hammerheads thus substituted at any of these positions showed an approximately 75-fold reduction in kcat whereas Km was unaffected. Hammerheads with modifications at G5 or G8 showed a significant reduction in magnesium binding affinity whereas modification at G12 had no effect. The results show that the three conserved G residues play crucial but different role sin hammerhead cleavage.
摘要
描述了含有修饰核苷O6-甲基鸟苷的寡核糖核苷酸的合成。通过使用2'-甲硅烷基保护的核苷亚磷酰胺、一种新的含O6-甲基鸟苷的合成子和温和的脱保护方法进行固相寡核糖核苷酸组装。O6-甲基鸟苷修饰的寡核苷酸用于研究锤头状核酶切割中保守残基G5、G8和G12的作用。在这些位置中的任何一个位置被取代的锤头状核酶的kcat降低了约75倍，而Km不受影响。在G5或G8处有修饰的锤头状核酶的镁结合亲和力显著降低，而在G12处的修饰没有影响。结果表明，这三个保守的G残基在锤头状核酶切割中起关键但不同的作用。
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