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 language | English (US) |
|---|---|
| Pages (from-to) | 27-44 |
| Number of pages | 18 |
| Journal | Somatic Cell Genetics |
| Volume | 4 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 1978 |
| Externally published | Yes |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Six complementation classes of conditionally lethal protein synthesis mutants of CHO cells selected by3H-amino acid
AU - Adair, Gerald M.
AU - Thompson, Larry H.
AU - Lindl, Patricia A.
PY - 1978/1
Y1 - 1978/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0017869953&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0017869953&partnerID=8YFLogxK
U2 - 10.1007/BF01546491
DO - 10.1007/BF01546491
M3 - Article
C2 - 628883
AN - SCOPUS:0017869953
VL - 4
SP - 27
EP - 44
JO - Somatic Cell and Molecular Genetics
JF - Somatic Cell and Molecular Genetics
SN - 0740-7750
IS - 1
ER -