Open AccessIdentification of important bases in a single‐stranded region (SSrC) of the hepatitis delta (δ) virus ribozyme
Author(s) -
KAWAKAMI Junji,
KUMAR P. K. R.,
SUH YoungAh,
NISHIKAWA Fumiko,
KAWAKAMI Keiko,
TAIRA Kazunari,
OHTSUKA Eiko,
NISHIKAWA Satoshi
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb18214.x
Subject(s) - hepatitis delta , ribozyme , virology , delta , identification (biology) , virus , mammalian cpeb3 ribozyme , biology , hepatitis b virus , physics , rna , genetics , gene , botany , astronomy
Models for the secondary structure of genomic and antigenomic self‐cleaving RNAs of human hepatitis delta (δ) virus (HDV) have been proposed by several groups. Our recent results support a pseudoknot structure and have allowed us to identify functionally important nucleotides in single‐stranded regions [nucleotides 726–731 (SSrA) and nucleotides 762–766 (SSrB)]. For the identification of the important residues in the remaining single‐stranded region, nucleotides 708–715 (SSrC), of the genomic HDV ribozyme, we made derivatives with a single‐base substitution in the SSrC region. To screen inactive mutants rapidly, we use a simplified in‐vitro selection method. Among the various base substitutions in mutants in the SSrC, U708A, C709(A/G/U) and G713C variants had less than 10% of the cleavage activity of the wild‐type SSrC (HDV86). By analyzing the self‐cleavage activities of various mutants, we determined the base requirements for SSrC as 5′‐(U/C/G)‐C‐N‐N‐(C/A/G)‐(G/A/U)‐N‐N‐3′.