Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1985 Sep;5(9):2381–2388. doi: 10.1128/mcb.5.9.2381

Amplification of the gene for histidyl-tRNA synthetase in histidinol-resistant Chinese hamster ovary cells.

F W Tsui, I L Andrulis, H Murialdo, L Siminovitch
PMCID: PMC366965  PMID: 2874482

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.

Full text

PDF

Images in this article

2384Image
on p.2384
2385Image
on p.2385
2385Image
on p.2385
2386Image
on p.2386
2386Image
on p.2386
2386Image
on p.2386
2387Image
on p.2387
2387Image
on p.2387

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adair G. M., Thompson L. H., Lindl P. A. Six complementation classes of conditionally lethal protein synthesis mutants of CHO cells selected by 3H-amino acid. Somatic Cell Genet. 1978 Jan;4(1):27–44. doi: 10.1007/BF01546491. [DOI] [PubMed] [Google Scholar]
  2. Andrulis I. L., Chiang C. S., Arfin S. M., Miner T. A., Hatfield G. W. Biochemical characterization of a mutant asparaginyl-tRNA synthetase from Chinese hamster ovary cells. J Biol Chem. 1978 Jan 10;253(1):58–62. [PubMed] [Google Scholar]
  3. Ashman C. R. Mutations in the structural genes of CHO cell histidyl-, valyl-, and leucyl-tRNA synthetases. Somatic Cell Genet. 1978 May;4(3):299–312. doi: 10.1007/BF01542844. [DOI] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. Cirullo R. E., Wasmuth J. J. Isolation of Chinese hamster ovary cells that overproduce asparaginyl-tRNA synthetase. Mol Cell Biol. 1984 Sep;4(9):1939–1941. doi: 10.1128/mcb.4.9.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Debatisse M., de Saint Vincent B. R., Buttin G. Expression of several amplified genes in an adenylate-deaminase overproducing variant of Chinese hamster fibroblasts. EMBO J. 1984 Dec 20;3(13):3123–3127. doi: 10.1002/j.1460-2075.1984.tb02268.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gerken S. C., Arfin S. M. Chinese hamster ovary cells resistant to borrelidin overproduce threonyl-tRNA synthetase. J Biol Chem. 1984 Jul 25;259(14):9202–9206. [PubMed] [Google Scholar]
  9. Gerken S. C., Arfin S. M. Threonyl-tRNA synthetase from Chinese hamster ovary cells is phosphorylated on serine. J Biol Chem. 1984 Sep 25;259(18):11160–11161. [PubMed] [Google Scholar]
  10. Kessler S. W. Cell membrane antigen isolation with the staphylococcal protein A-antibody adsorbent. J Immunol. 1976 Nov;117(5 Pt 1):1482–1490. [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lehrach H., Frischauf A. M., Hanahan D., Wozney J., Fuller F., Boedtker H. Construction and characterization of pro alpha 1 collagen complementary deoxyribonucleic acid clones. Biochemistry. 1979 Jul 10;18(14):3146–3152. doi: 10.2196/47873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mathews M. B., Bernstein R. M. Myositis autoantibody inhibits histidyl-tRNA synthetase: a model for autoimmunity. Nature. 1983 Jul 14;304(5922):177–179. doi: 10.1038/304177a0. [DOI] [PubMed] [Google Scholar]
  15. Padgett R. A., Wahl G. M., Coleman P. F., Stark G. R. N-(Phosphonacetyl)-L-aspartate-resistant hamster cells overaccumulate a single mRNA coding for the multifunctional protein that catalyzes the first steps of UMP synthesis. J Biol Chem. 1979 Feb 10;254(3):974–980. [PubMed] [Google Scholar]
  16. Schimke R. T. Gene amplification in cultured animal cells. Cell. 1984 Jul;37(3):705–713. doi: 10.1016/0092-8674(84)90406-9. [DOI] [PubMed] [Google Scholar]
  17. Scornik O. A., Ledbetter M. L., Malter J. S. Role of aminoacylation of histidyl-tRNA in the regulation of protein degradation in Chinese hamster ovary cells. J Biol Chem. 1980 Jul 10;255(13):6322–6329. [PubMed] [Google Scholar]
  18. Stanners C. P., Eliceiri G. L., Green H. Two types of ribosome in mouse-hamster hybrid cells. Nat New Biol. 1971 Mar 10;230(10):52–54. doi: 10.1038/newbio230052a0. [DOI] [PubMed] [Google Scholar]
  19. Stark G. R., Wahl G. M. Gene amplification. Annu Rev Biochem. 1984;53:447–491. doi: 10.1146/annurev.bi.53.070184.002311. [DOI] [PubMed] [Google Scholar]
  20. Thompson L. H., Harkins J. L., Stanners C. P. A mammalian cell mutant with a temperature-sensitive leucyl-transfer RNA synthetase. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3094–3098. doi: 10.1073/pnas.70.11.3094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thompson L. H., Lofgren D. J., Adair G. M. Evidence for structural gene alterations affecting aminoacyl-tRNA synthetases in CHO cell mutants and revertants. Somatic Cell Genet. 1978 Jul;4(4):423–435. doi: 10.1007/BF01538864. [DOI] [PubMed] [Google Scholar]
  22. Thompson L. H., Stanners C. P., Siminovitch L. Selection by [3H] amino acids of CHO-cell mutants with altered leucyl- and asparagyl-transfer RNA synthetases. Somatic Cell Genet. 1975 Apr;1(2):187–208. doi: 10.1007/BF01538547. [DOI] [PubMed] [Google Scholar]
  23. Wahl G. M., Padgett R. A., Stark G. R. Gene amplification causes overproduction of the first three enzymes of UMP synthesis in N-(phosphonacetyl)-L-aspartate-resistant hamster cells. J Biol Chem. 1979 Sep 10;254(17):8679–8689. [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES