A new method for identifying the amino acid attached to a particular RNA in the cell

Tsutomu Suzuki; Takuya Ueda; Kimitsuna Watanabe

doi:10.1016/0014-5793(96)00107-X

A new method for identifying the amino acid attached to a particular RNA in the cell

Tsutomu Suzuki, Takuya Ueda, Kimitsuna Watanabe^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

16 Citations (Scopus)

Abstract

To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [¹⁴C]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The ¹⁴C-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.

Original language	English
Pages (from-to)	195-198
Number of pages	4
Journal	FEBS Letters
Volume	381
Issue number	3
DOIs	https://doi.org/10.1016/0014-5793(96)00107-X
Publication status	Published - 1996 Mar 4
Externally published	Yes

Keywords

Amino acid
Aminoacyl-tRNA (in vivo)
Candida
Hybridization
N-Acetylation

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(96)00107-X

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abstract = "To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [14C]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The 14C-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.",

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AU - Suzuki, Tsutomu

AU - Ueda, Takuya

AU - Watanabe, Kimitsuna

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Y1 - 1996/3/4

N2 - To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [14C]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The 14C-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.

AB - To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [14C]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The 14C-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.

KW - Amino acid

KW - Aminoacyl-tRNA (in vivo)

KW - Candida

KW - Hybridization

KW - N-Acetylation

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A new method for identifying the amino acid attached to a particular RNA in the cell

Abstract

Keywords

ASJC Scopus subject areas

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