Amplification of the gene for histidyl-tRNA synthetase in histidinol-resistant Chinese hamster ovary cells.
Open Access
- 1 September 1985
- journal article
- research article
- Published by Taylor & Francis in Molecular and Cellular Biology
- Vol. 5 (9) , 2381-2388
- https://doi.org/10.1128/mcb.5.9.2381
Abstract
Histidinol-resistant (HisOHR) mutants with up to a 30-fold increase in histidyl-tRNA synthetase activity have been isolated by stepwise adaptation of wild-type Chinese hamster ovary (CHO) cells to increasing amounts of histidinol in the medium. Immunoprecipitation of [35S]methionine-labeled cell lysates with antibodies to histidyl-tRNA synthetase showed increased synthesis of the enzyme in histidinol-resistant cells. The histidinol-resistant cell lines had an increase in translatable polyadenylated mRNA for histidyl-tRNA synthetase. A cDNA for CHO histidyl-tRNA synthetase has been cloned, using these histidyl-tRNA synthetase-overproducing mutants as the source of mRNA. Southern blot analysis of wild-type and histidinol-resistant cells with this cDNA showed that the histidyl-tRNA synthetase DNA bands were amplified in the resistant cells. These HisOHR cells owed their resistance to histidinol to amplification of the gene for histidyl-tRNA synthetase.This publication has 24 references indexed in Scilit:
- Evidence for structural gene alterations affecting aminoacyl-tRNA synthetases in CHO cell mutants and revertantsSomatic Cell and Molecular Genetics, 1978
- Mutations in the structural genes of CHO cell histidyl-, valyl-, and leucyl-tRNA synthetasesSomatic Cell and Molecular Genetics, 1978
- BIOCHEMICAL CHARACTERIZATION OF A MUTANT ASPARAGINYL TRANSFER-RNA SYNTHETASE FROM CHINESE-HAMSTER OVARY CELLS1978
- Six complementation classes of conditionally lethal protein synthesis mutants of CHO cells selected by3H-amino acidSomatic Cell and Molecular Genetics, 1978
- Cell Membrane Antigen Isolation with the Staphylococcal Protein A-Antibody AdsorbentThe Journal of Immunology, 1976
- Selection by [3H] amino acids of CHO-cell mutants with altered leucyl- and asparagyl-transfer RNA synthetasesSomatic Cell and Molecular Genetics, 1975
- A Film Detection Method for Tritium‐Labelled Proteins and Nucleic Acids in Polyacrylamide GelsEuropean Journal of Biochemistry, 1974
- A Mammalian Cell Mutant with a Temperature-Sensitive Leucyl-Transfer RNA SynthetaseProceedings of the National Academy of Sciences, 1973
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951