Role of the extra G-C pair at the end of the acceptor stem of tRNAHb in aminoacylation

Hyouta Himeno, Tsunemi Hasegawa, Takuya Ueda, Kimitsuna Watanabe, Kin Ichiro Miura, Mikio Shimizu

Research output: Contribution to journalArticle

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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 aainoacylation, 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 systm. 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.

Original languageEnglish
Pages (from-to)7855-7863
Number of pages9
JournalNucleic acids research
Volume17
Issue number19
DOIs
Publication statusPublished - 1989 Oct 11
Externally publishedYes

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Aminoacylation
Transfer RNA
Histidine-tRNA Ligase
Histidine
Transfer RNA Aminoacylation
Base Pairing
Nucleotides
Escherichia coli

ASJC Scopus subject areas

  • Genetics

Cite this

Himeno, H., Hasegawa, T., Ueda, T., Watanabe, K., Miura, K. I., & Shimizu, M. (1989). Role of the extra G-C pair at the end of the acceptor stem of tRNAHb in aminoacylation. Nucleic acids research, 17(19), 7855-7863. https://doi.org/10.1093/nar/17.19.7855

Role of the extra G-C pair at the end of the acceptor stem of tRNAHb in aminoacylation. / Himeno, Hyouta; Hasegawa, Tsunemi; Ueda, Takuya; Watanabe, Kimitsuna; Miura, Kin Ichiro; Shimizu, Mikio.

In: Nucleic acids research, Vol. 17, No. 19, 11.10.1989, p. 7855-7863.

Research output: Contribution to journalArticle

Himeno, H, Hasegawa, T, Ueda, T, Watanabe, K, Miura, KI & Shimizu, M 1989, 'Role of the extra G-C pair at the end of the acceptor stem of tRNAHb in aminoacylation', Nucleic acids research, vol. 17, no. 19, pp. 7855-7863. https://doi.org/10.1093/nar/17.19.7855
Himeno, Hyouta ; Hasegawa, Tsunemi ; Ueda, Takuya ; Watanabe, Kimitsuna ; Miura, Kin Ichiro ; Shimizu, Mikio. / Role of the extra G-C pair at the end of the acceptor stem of tRNAHb in aminoacylation. In: Nucleic acids research. 1989 ; Vol. 17, No. 19. pp. 7855-7863.
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