Conversion of aminoacylation specificity from tRNATyr to tRNASer in vitro

Hyouta Himeno, Tsunemi Hasegawa, Takuya Ueda, Kimitsuna Watanabe, Mikio Shimizu

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

The discrimination mechanism between tRNASer and tRNATyr was studied using various in vitro transcripts of E. coli tRNATyr variants. The insertion of only two nucleotldes into the variable stem of tRNATyr generates serine charging activity. The acceptor activities of some of the tRNATyr mutants with insertions in the long variable arm were enhanced by changes in nucleotides at positions 9 and/or 20B, which are possible elements for dictating the orientation of the long variable arm. These findings suggest that the long variable arm is involved in recognition by seryl-tRNA synthetase in spite of sequence and length variations shown within tRNASer isoacceptors, and eventually serves as a determinant for selection from other tRNA species. Changing the anticodon from GUA to the serine anticodon GGA resulted in a marked decrease in tyrosine charging activity, but this mutant did not show any serine charging activity. The discriminator base, the fourth base from the 3′ end of tRNA, was also Important for aminoacylation with tyrosine. Complete specificity change in vitro was facilitated by insertion of three nucleotides into the variable arm plus two nucleotide changes at positions 9 and 73.

Original languageEnglish
Pages (from-to)6815-6819
Number of pages5
JournalNucleic acids research
Volume18
Issue number23
DOIs
Publication statusPublished - 1990 Dec 1
Externally publishedYes

Fingerprint

RNA, Transfer, Tyr
RNA, Transfer, Ser
Aminoacylation
Serine
Anticodon
Nucleotides
Transfer RNA
Tyrosine
Serine-tRNA Ligase
Escherichia coli
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics

Cite this

Conversion of aminoacylation specificity from tRNATyr to tRNASer in vitro. / Himeno, Hyouta; Hasegawa, Tsunemi; Ueda, Takuya; Watanabe, Kimitsuna; Shimizu, Mikio.

In: Nucleic acids research, Vol. 18, No. 23, 01.12.1990, p. 6815-6819.

Research output: Contribution to journalArticle

Himeno, Hyouta ; Hasegawa, Tsunemi ; Ueda, Takuya ; Watanabe, Kimitsuna ; Shimizu, Mikio. / Conversion of aminoacylation specificity from tRNATyr to tRNASer in vitro. In: Nucleic acids research. 1990 ; Vol. 18, No. 23. pp. 6815-6819.
@article{5264607bf51b497091e863dfcaa0ed65,
title = "Conversion of aminoacylation specificity from tRNATyr to tRNASer in vitro",
abstract = "The discrimination mechanism between tRNASer and tRNATyr was studied using various in vitro transcripts of E. coli tRNATyr variants. The insertion of only two nucleotldes into the variable stem of tRNATyr generates serine charging activity. The acceptor activities of some of the tRNATyr mutants with insertions in the long variable arm were enhanced by changes in nucleotides at positions 9 and/or 20B, which are possible elements for dictating the orientation of the long variable arm. These findings suggest that the long variable arm is involved in recognition by seryl-tRNA synthetase in spite of sequence and length variations shown within tRNASer isoacceptors, and eventually serves as a determinant for selection from other tRNA species. Changing the anticodon from GUA to the serine anticodon GGA resulted in a marked decrease in tyrosine charging activity, but this mutant did not show any serine charging activity. The discriminator base, the fourth base from the 3′ end of tRNA, was also Important for aminoacylation with tyrosine. Complete specificity change in vitro was facilitated by insertion of three nucleotides into the variable arm plus two nucleotide changes at positions 9 and 73.",
author = "Hyouta Himeno and Tsunemi Hasegawa and Takuya Ueda and Kimitsuna Watanabe and Mikio Shimizu",
year = "1990",
month = "12",
day = "1",
doi = "10.1093/nar/18.23.6815",
language = "English",
volume = "18",
pages = "6815--6819",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "23",

}

TY - JOUR

T1 - Conversion of aminoacylation specificity from tRNATyr to tRNASer in vitro

AU - Himeno, Hyouta

AU - Hasegawa, Tsunemi

AU - Ueda, Takuya

AU - Watanabe, Kimitsuna

AU - Shimizu, Mikio

PY - 1990/12/1

Y1 - 1990/12/1

N2 - The discrimination mechanism between tRNASer and tRNATyr was studied using various in vitro transcripts of E. coli tRNATyr variants. The insertion of only two nucleotldes into the variable stem of tRNATyr generates serine charging activity. The acceptor activities of some of the tRNATyr mutants with insertions in the long variable arm were enhanced by changes in nucleotides at positions 9 and/or 20B, which are possible elements for dictating the orientation of the long variable arm. These findings suggest that the long variable arm is involved in recognition by seryl-tRNA synthetase in spite of sequence and length variations shown within tRNASer isoacceptors, and eventually serves as a determinant for selection from other tRNA species. Changing the anticodon from GUA to the serine anticodon GGA resulted in a marked decrease in tyrosine charging activity, but this mutant did not show any serine charging activity. The discriminator base, the fourth base from the 3′ end of tRNA, was also Important for aminoacylation with tyrosine. Complete specificity change in vitro was facilitated by insertion of three nucleotides into the variable arm plus two nucleotide changes at positions 9 and 73.

AB - The discrimination mechanism between tRNASer and tRNATyr was studied using various in vitro transcripts of E. coli tRNATyr variants. The insertion of only two nucleotldes into the variable stem of tRNATyr generates serine charging activity. The acceptor activities of some of the tRNATyr mutants with insertions in the long variable arm were enhanced by changes in nucleotides at positions 9 and/or 20B, which are possible elements for dictating the orientation of the long variable arm. These findings suggest that the long variable arm is involved in recognition by seryl-tRNA synthetase in spite of sequence and length variations shown within tRNASer isoacceptors, and eventually serves as a determinant for selection from other tRNA species. Changing the anticodon from GUA to the serine anticodon GGA resulted in a marked decrease in tyrosine charging activity, but this mutant did not show any serine charging activity. The discriminator base, the fourth base from the 3′ end of tRNA, was also Important for aminoacylation with tyrosine. Complete specificity change in vitro was facilitated by insertion of three nucleotides into the variable arm plus two nucleotide changes at positions 9 and 73.

UR - http://www.scopus.com/inward/record.url?scp=0025651675&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025651675&partnerID=8YFLogxK

U2 - 10.1093/nar/18.23.6815

DO - 10.1093/nar/18.23.6815

M3 - Article

VL - 18

SP - 6815

EP - 6819

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 23

ER -