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SYSTEMATIC SUBSTITUTION OF INDIVIDUAL BASES IN 2 IMPORTANT SINGLE-STRANDED REGIONS OF THE HDV RIBOZYME FOR EVALUATION OF THE ROLE OF SPECIFIC BASES

被引：24
作者：
SUH, YA
KUMAR, PKR
KAWAKAMI, J
NISHIKAWA, F
TAIRA, K
NISHIKAWA, S
机构：
[1] MINIST INT TRADE & IND,AGCY IND SCI & TECHNOL,NATL INST BIOSCI & HUMAN TECHNOL,TSUKUBA 305,JAPAN
[2] HOKKAIDO UNIV,DIV PHARMACEUT SCI,SAPPORO,HOKKAIDO 060,JAPAN
来源：
FEBS LETTERS | 1993年 / 326卷 / 1-3期
关键词：
HDV RIBOZYME; SELF-CLEAVAGE ACTIVITY; PSEUDOKNOT STRUCTURE; SINGLE-STRANDED REGION; POINT MUTATION;
D O I：
10.1016/0014-5793(93)81782-U
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：
To elucidate the role of specific bases in the self-cleavage activity of the human hepatitis delta virus (HDV) riboxyme, systematic substitutions of individual bases in two important single-stranded regions [between nucleotides 726-731 (SSrA region) and 762-766 (SSrB region)] were carried out by oligonucleotide-directed point mutagenesis. Among the mutants obtained, 12 mutants (G726 variants, G727A, G727C, G728C, G762A, G762C, C763 variants and A766C) could not tolerate the respective base-substitutions and self-cleavage activities were reduced to very low levels (10%), suggesting a requirement of the respective bases. In particular, G726 in the SSrA region and C763 in the SSrB region were found to be essential for the ribozyme activity. We could determine the preferred sequences, 5'-G-G-(G/A/U)-N-(A/U/G)-Pu-3' for SSrA and 5'-(G/U)-C-N-(A/G/U)-A-3' for SSrB regions, respectively.
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页码：158 / 162
页数：5
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