CHO cell mutants for arginyl-, asparagyl-, glutaminyl-, histidyl- and methionyl-transfer RNA synthetases: Identification and initial characterization

Larry H. Thompson, Don J. Lofgren, Gerald M. Adair

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

A number of conditional lethal mutants of CHO cells that are defective in protein synthesis have been characterized with respect to their biochemical lesions. A defective aminoacyl-tRNA synthetase appears to be the basis of each mutant phenotype. In each strain, the specific activity in vitro of the synthetase cognate for one of the following amino acids was substantially reduced: arginine, asparagine, glutamine, histidine or methionine. One mutant, Arg-1, gave no detectable arginyl-tRNA synthetase activity, suggesting that it contains an altered enzyme that is unstable in vitro. Most of the mutants correspondingly exhibited impaired aminoacylation in vivo under nonpermissive conditions. However, two mutants, Arg-1 and His-1, appeared to have normal levels of acylated tRNA under the nonpermissive conditions which inhibited protein synthesis to approximately 50% and 10%, respectively. The expression of each mutant's phenotype, measured by rates of protein synthesis or growth, was a function of temperature and/or the concentration of amino acid cognate for the synthetase found to be deficient in vitro. The properties of these mutants make them applicable to diverse problems related to translation in mammalian cells.

Original languageEnglish (US)
Pages (from-to)157-168
Number of pages12
JournalCell
Volume11
Issue number1
DOIs
StatePublished - 1977
Externally publishedYes

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Amino Acyl-tRNA Synthetases
CHO Cells
Ligases
Arginine-tRNA Ligase
Aminoacylation
Phenotype
Amino Acids
Proteins
Asparagine
Transfer RNA
Glutamine
Histidine
Methionine
Arginine
Cells
Temperature
Enzymes
Growth
In Vitro Techniques

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

CHO cell mutants for arginyl-, asparagyl-, glutaminyl-, histidyl- and methionyl-transfer RNA synthetases : Identification and initial characterization. / Thompson, Larry H.; Lofgren, Don J.; Adair, Gerald M.

In: Cell, Vol. 11, No. 1, 1977, p. 157-168.

Research output: Contribution to journalArticle

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abstract = "A number of conditional lethal mutants of CHO cells that are defective in protein synthesis have been characterized with respect to their biochemical lesions. A defective aminoacyl-tRNA synthetase appears to be the basis of each mutant phenotype. In each strain, the specific activity in vitro of the synthetase cognate for one of the following amino acids was substantially reduced: arginine, asparagine, glutamine, histidine or methionine. One mutant, Arg-1, gave no detectable arginyl-tRNA synthetase activity, suggesting that it contains an altered enzyme that is unstable in vitro. Most of the mutants correspondingly exhibited impaired aminoacylation in vivo under nonpermissive conditions. However, two mutants, Arg-1 and His-1, appeared to have normal levels of acylated tRNA under the nonpermissive conditions which inhibited protein synthesis to approximately 50{\%} and 10{\%}, respectively. The expression of each mutant's phenotype, measured by rates of protein synthesis or growth, was a function of temperature and/or the concentration of amino acid cognate for the synthetase found to be deficient in vitro. The properties of these mutants make them applicable to diverse problems related to translation in mammalian cells.",
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