CHEMICAL PROBING STUDIES OF VARIANTS OF THE GENOMIC HEPATITIS-DELTA VIRUS RIBOZYME BY PRIMER EXTENSION ANALYSIS

被引：37

作者：

KUMAR, PKR ^{[1
]}

TAIRA, K ^{[1
]}

NISHIKAWA, S ^{[1
]}

机构：

[1] MINIST INT TRADE & IND, AGCY IND SCI & TECHNOL, NATL INST BIOSCI & HUMAN TECHNOL, TSUKUBA, IBARAKI 305, JAPAN

来源：

BIOCHEMISTRY | 1994年 / 33卷 / 02期

关键词：

D O I：

10.1021/bi00168a025

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have investigated in detail the higher order structure of the genomic hepatitis delta virus (HDV) ribozyme using various base-specific chemical probes under native, semi-denaturing, and denaturing conditions. The bases of the HDV ribozyme were probed by treatment with dimethyl sulfate [which reacts with A (at N1) and C (at N3)] and a carbodiimide [which reacts with U (at N3) and G (at N1)]. In addition, for probing G residues (at N7), RNA samples were treated with NaBH4 and aniline after modification by treatment with dimethyl sulfate. The sites of modified positions were identified by primer extension analysis with reverse transcriptase. In general, our results are consistent with the proposed pseudoknot model of secondary structure, a model that is based on data from ribonucleolytic cleavage experiments. Our results provide clues to the identification of interacting bases in the HDV ribozyme. Furthermore, using this method we identified local conformational changes in several stem variants.

引用

页码：583 / 592

页数：10

共 27 条

[1] SECONDARY STRUCTURE OF THE SELF-CLEAVING RNA OF HEPATITIS DELTA-VIRUS - APPLICATIONS TO CATALYTIC RNA DESIGN [J].