Six complementation classes of conditionally lethal protein synthesis mutants of CHO cells selected by3H-amino acid

Gerald M. Adair, Larry H. Thompson, Patricia A. Lindl

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Using a tritiated amino acid suicide procedure designed specifically to select conditional protein synthesis mutants, we have isolated and characterized a large number of such mutants of Chinese hamster ovary cells. All of the mutants are genetically stable and behave as recessives in somatic cell hybrids. Most of the new mutants are phenotypically dependent on the concentration of a specific amino acid as well as on temperature. In addition to identifying many additional leucyl- and asparagyl-tRNA synthetase mutants, complementation analysis has distinguished four new genetic classes representing methionine-, glutamine-, histidine-, and arginine-dependent mutants. Biochemical characterization of representative mutants from each of these six classes has identified the primary lesions as being defective aminoacyl-tRNA synthetases. Our selection results further demonstrate the high specificity of the3H-amino acid procedure for isolating protein synthesis mutants. Reconstruction experiments performed with two representative mutants indicated a selection efficiency of approximately 10% under standard conditions.

Original languageEnglish (US)
Pages (from-to)27-44
Number of pages18
JournalSomatic Cell Genetics
Volume4
Issue number1
DOIs
StatePublished - Jan 1978
Externally publishedYes

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CHO Cells
Mutant Proteins
Amino Acyl-tRNA Synthetases
Amino Acids
Hybrid Cells
Cricetulus
Glutamine
Histidine
Methionine
Suicide
Arginine
Ovary
Temperature

ASJC Scopus subject areas

  • Genetics
  • Medicine(all)

Cite this

Six complementation classes of conditionally lethal protein synthesis mutants of CHO cells selected by3H-amino acid. / Adair, Gerald M.; Thompson, Larry H.; Lindl, Patricia A.

In: Somatic Cell Genetics, Vol. 4, No. 1, 01.1978, p. 27-44.

Research output: Contribution to journalArticle

Adair, Gerald M. ; Thompson, Larry H. ; Lindl, Patricia A. / Six complementation classes of conditionally lethal protein synthesis mutants of CHO cells selected by3H-amino acid. In: Somatic Cell Genetics. 1978 ; Vol. 4, No. 1. pp. 27-44.
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