title: Role of the extra G-C pair at the end of the acceptor stem of tRNA(His) in aminoacylation.
abstract: All sequenced histidine tRNAs have one additional nucleotide at the 5' end compared with other tRNA species. To investigate the role of this unique structure in aminoacylation, we constructed in vitro transcripts corresponding to the E. coli histidine tRNA sequence and its variants at the G-1-C73 base pair, by using T7 RNA polymerase transcription system. A transcript having a wild-type sequence with no modified bases was a good substrate for histidyl-tRNA synthetase (HisRS), and aminoacylation activity was affected by introduction of a triphosphate at the 5' terminus. Base replacements at position 73 caused a marked decrease of Vmax, and deletion and substitution of the G-1 had a remarkable effect on the aminoacylation. A mutant having an A-1-U73 pair was also not a good substrate for HisRS. Comparison among G-1-deficient mutants showed that A was preferable rather than C as the base at position 73. These data demonstrate that the set of the G-1-C73 pair at the end of the acceptor stem of histidine tRNA is crucial for the catalytic process of aminoacylation.
authors.affiliation: Institute of Space and Astronautical Science, Kanagawa, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Himeno
authors.forename: T
authors.initials: T
authors.lastname: Hasegawa
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
authors.forename: K
authors.initials: K
authors.lastname: Miura
authors.forename: M
authors.initials: M
authors.lastname: Shimizu
mesh_terms: D000215:Acylation; D001482:Base Composition; D001483:Base Sequence; D003596:Cytosine; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D006147:Guanine; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D012354:RNA, Transfer, His / Q000235:genetics*; D015183:Restriction Mapping; D013604:T-Phages / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: D012345:RNA, Transfer, Amino Acid-Specific; D012354:RNA, Transfer, His; D006147:Guanine; D003596:Cytosine; D012321:DNA-Directed RNA Polymerases