Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 May;85(10):3499–3503. doi: 10.1073/pnas.85.10.3499

Spectrum of spontaneous mutation at the APRT locus of Chinese hamster ovary cells: an analysis at the DNA sequence level.

P J de Jong 1, A J Grosovsky 1, B W Glickman 1
PMCID: PMC280239  PMID: 3368459

Abstract

The spectrum of spontaneous mutation of an endogenous mammalian cell gene has been determined at the DNA sequence level. Thirty independent spontaneous APRT- mutations were cloned and subsequently completely sequenced. Twenty-seven contained single base substitutions. Of these, 22 were G.C to A.T transitions, suggesting a major role for the deamination of cytosine in spontaneous mutagenesis of Chinese hamster ovary cells. The remaining mutants included a tandem double substitution, a -1 frameshift, and a 17-base-pair deletion flanked by a 2-base-pair direct repeat. Many of the independently recovered mutants were clustered at sites of multiple occurrence (hot spots). One site accounted for greater than 25% of all independently recovered events. Mutations were generally located within the coding sequence, although two mutations occurred within the consensus sequence for a 3' splice site.

Full text

PDF
3499

Selected References

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

  1. Albertini A. M., Hofer M., Calos M. P., Miller J. H. On the formation of spontaneous deletions: the importance of short sequence homologies in the generation of large deletions. Cell. 1982 Jun;29(2):319–328. doi: 10.1016/0092-8674(82)90148-9. [DOI] [PubMed] [Google Scholar]
  2. Aust A. E., Drinkwater N. R., Debien K., Maher V. M., McCormick J. J. Comparison of the frequency of diphtheria toxin and thioguanine resistance induced by a series of carcinogens to analyze their mutational specificities in diploid human fibroblasts. Mutat Res. 1984 Jan;125(1):95–104. doi: 10.1016/0027-5107(84)90036-8. [DOI] [PubMed] [Google Scholar]
  3. Benzer S. FINE STRUCTURE OF A GENETIC REGION IN BACTERIOPHAGE. Proc Natl Acad Sci U S A. 1955 Jun 15;41(6):344–354. doi: 10.1073/pnas.41.6.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bird A. P. DNA methylation and the frequency of CpG in animal DNA. Nucleic Acids Res. 1980 Apr 11;8(7):1499–1504. doi: 10.1093/nar/8.7.1499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradley W. E., Letovanec D. High-frequency nonrandom mutational event at the adenine phosphoribosyltransferase (aprt) locus of sib-selected CHO variants heterozygous for aprt. Somatic Cell Genet. 1982 Jan;8(1):51–66. doi: 10.1007/BF01538650. [DOI] [PubMed] [Google Scholar]
  6. Calos M. P., Lebkowski J. S., Botchan M. R. High mutation frequency in DNA transfected into mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3015–3019. doi: 10.1073/pnas.80.10.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chasin L. A. Mutations affecting adenine phosphoribosyl transferase activity in Chinese hamster cells. Cell. 1974 May;2(1):37–41. doi: 10.1016/0092-8674(74)90006-3. [DOI] [PubMed] [Google Scholar]
  8. Coulondre C., Miller J. H., Farabaugh P. J., Gilbert W. Molecular basis of base substitution hotspots in Escherichia coli. Nature. 1978 Aug 24;274(5673):775–780. doi: 10.1038/274775a0. [DOI] [PubMed] [Google Scholar]
  9. Duncan B. K., Miller J. H. Mutagenic deamination of cytosine residues in DNA. Nature. 1980 Oct 9;287(5782):560–561. doi: 10.1038/287560a0. [DOI] [PubMed] [Google Scholar]
  10. Dush M. K., Sikela J. M., Khan S. A., Tischfield J. A., Stambrook P. J. Nucleotide sequence and organization of the mouse adenine phosphoribosyltransferase gene: presence of a coding region common to animal and bacterial phosphoribosyltransferases that has a variable intron/exon arrangement. Proc Natl Acad Sci U S A. 1985 May;82(9):2731–2735. doi: 10.1073/pnas.82.9.2731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farabaugh P. J., Schmeissner U., Hofer M., Miller J. H. Genetic studies of the lac repressor. VII. On the molecular nature of spontaneous hotspots in the lacI gene of Escherichia coli. J Mol Biol. 1978 Dec 25;126(4):847–857. doi: 10.1016/0022-2836(78)90023-2. [DOI] [PubMed] [Google Scholar]
  12. Fix D. F., Glickman B. W. Asymmetric cytosine deamination revealed by spontaneous mutational specificity in an Ung- strain of Escherichia coli. Mol Gen Genet. 1987 Aug;209(1):78–82. doi: 10.1007/BF00329839. [DOI] [PubMed] [Google Scholar]
  13. Fix D. F., Glickman B. W. Differential enhancement of spontaneous transition mutations in the lacI gene of an Ung- strain of Escherichia coli. Mutat Res. 1986 Oct;175(2):41–45. doi: 10.1016/0165-7992(86)90123-5. [DOI] [PubMed] [Google Scholar]
  14. Glickman B. W., Ripley L. S. Structural intermediates of deletion mutagenesis: a role for palindromic DNA. Proc Natl Acad Sci U S A. 1984 Jan;81(2):512–516. doi: 10.1073/pnas.81.2.512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grosovsky A. J., Drobetsky E. A., deJong P. J., Glickman B. W. Southern analysis of genomic alterations in gamma-ray-induced aprt- hamster cell mutants. Genetics. 1986 Jun;113(2):405–415. doi: 10.1093/genetics/113.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones G. E., Sargent P. A. Mutants of cultured chinese hamster cells deficient in adenine phosphoribosyl transferase. Cell. 1974 May;2(1):43–54. doi: 10.1016/0092-8674(74)90007-5. [DOI] [PubMed] [Google Scholar]
  17. Kunkel T. A., Alexander P. S. The base substitution fidelity of eucaryotic DNA polymerases. Mispairing frequencies, site preferences, insertion preferences, and base substitution by dislocation. J Biol Chem. 1986 Jan 5;261(1):160–166. [PubMed] [Google Scholar]
  18. Lebkowski J. S., DuBridge R. B., Antell E. A., Greisen K. S., Calos M. P. Transfected DNA is mutated in monkey, mouse, and human cells. Mol Cell Biol. 1984 Oct;4(10):1951–1960. doi: 10.1128/mcb.4.10.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Leong P. M., Thilly W. G., Morgenthaler S. Variance estimation in single-cell mutation assays: comparison to experimental observations in human lymphoblasts at 4 gene loci. Mutat Res. 1985 Jun-Jul;150(1-2):403–410. doi: 10.1016/0027-5107(85)90137-x. [DOI] [PubMed] [Google Scholar]
  20. Levinson A., Silver D., Seed B. Minimal size plasmids containing an M13 origin for production of single-strand transducing particles. J Mol Appl Genet. 1984;2(6):507–517. [PubMed] [Google Scholar]
  21. Li W. H., Wu C. I., Luo C. C. Nonrandomness of point mutation as reflected in nucleotide substitutions in pseudogenes and its evolutionary implications. J Mol Evol. 1984;21(1):58–71. doi: 10.1007/BF02100628. [DOI] [PubMed] [Google Scholar]
  22. Lindahl T., Nyberg B. Heat-induced deamination of cytosine residues in deoxyribonucleic acid. Biochemistry. 1974 Jul 30;13(16):3405–3410. doi: 10.1021/bi00713a035. [DOI] [PubMed] [Google Scholar]
  23. Meuth M., Arrand J. E. Alterations of gene structure in ethyl methane sulfonate-induced mutants of mammalian cells. Mol Cell Biol. 1982 Nov;2(11):1459–1462. doi: 10.1128/mcb.2.11.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miller J. H. Mutagenic specificity of ultraviolet light. J Mol Biol. 1985 Mar 5;182(1):45–65. doi: 10.1016/0022-2836(85)90026-9. [DOI] [PubMed] [Google Scholar]
  25. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nalbantoglu J., Goncalves O., Meuth M. Structure of mutant alleles at the aprt locus of Chinese hamster ovary cells. J Mol Biol. 1983 Jul 5;167(3):575–594. doi: 10.1016/s0022-2836(83)80099-0. [DOI] [PubMed] [Google Scholar]
  27. Nalbantoglu J., Hartley D., Phear G., Tear G., Meuth M. Spontaneous deletion formation at the aprt locus of hamster cells: the presence of short sequence homologies and dyad symmetries at deletion termini. EMBO J. 1986 Jun;5(6):1199–1204. doi: 10.1002/j.1460-2075.1986.tb04347.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nalbantoglu J., Phear G. A., Meuth M. Nucleotide sequence of hamster adenine phosphoribosyl transferase (aprt) gene. Nucleic Acids Res. 1986 Feb 25;14(4):1914–1914. doi: 10.1093/nar/14.4.1914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Razzaque A., Chakrabarti S., Joffee S., Seidman M. Mutagenesis of a shuttle vector plasmid in mammalian cells. Mol Cell Biol. 1984 Mar;4(3):435–441. doi: 10.1128/mcb.4.3.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Razzaque A., Mizusawa H., Seidman M. M. Rearrangement and mutagenesis of a shuttle vector plasmid after passage in mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3010–3014. doi: 10.1073/pnas.80.10.3010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reed R., Maniatis T. Intron sequences involved in lariat formation during pre-mRNA splicing. Cell. 1985 May;41(1):95–105. doi: 10.1016/0092-8674(85)90064-9. [DOI] [PubMed] [Google Scholar]
  32. Schaaper R. M., Danforth B. N., Glickman B. W. Mechanisms of spontaneous mutagenesis: an analysis of the spectrum of spontaneous mutation in the Escherichia coli lacI gene. J Mol Biol. 1986 May 20;189(2):273–284. doi: 10.1016/0022-2836(86)90509-7. [DOI] [PubMed] [Google Scholar]
  33. Seed B. Purification of genomic sequences from bacteriophage libraries by recombination and selection in vivo. Nucleic Acids Res. 1983 Apr 25;11(8):2427–2445. doi: 10.1093/nar/11.8.2427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Seidman M. M., Dixon K., Razzaque A., Zagursky R. J., Berman M. L. A shuttle vector plasmid for studying carcinogen-induced point mutations in mammalian cells. Gene. 1985;38(1-3):233–237. doi: 10.1016/0378-1119(85)90222-7. [DOI] [PubMed] [Google Scholar]
  35. Selker E. U., Stevens J. N. DNA methylation at asymmetric sites is associated with numerous transition mutations. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8114–8118. doi: 10.1073/pnas.82.23.8114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Simon A. E., Taylor M. W., Bradley W. E., Thompson L. H. Model involving gene inactivation in the generation of autosomal recessive mutants in mammalian cells in culture. Mol Cell Biol. 1982 Sep;2(9):1126–1133. doi: 10.1128/mcb.2.9.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Streisinger G., Okada Y., Emrich J., Newton J., Tsugita A., Terzaghi E., Inouye M. Frameshift mutations and the genetic code. This paper is dedicated to Professor Theodosius Dobzhansky on the occasion of his 66th birthday. Cold Spring Harb Symp Quant Biol. 1966;31:77–84. doi: 10.1101/sqb.1966.031.01.014. [DOI] [PubMed] [Google Scholar]
  38. Thacker J. The molecular nature of mutations in cultured mammalian cells: a review. Mutat Res. 1985 Jun-Jul;150(1-2):431–442. doi: 10.1016/0027-5107(85)90140-x. [DOI] [PubMed] [Google Scholar]
  39. Tindall K. R., Stankowski L. F., Jr, Machanoff R., Hsie A. W. Detection of deletion mutations in pSV2gpt-transformed cells. Mol Cell Biol. 1984 Jul;4(7):1411–1415. doi: 10.1128/mcb.4.7.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wu C. I., Maeda N. Inequality in mutation rates of the two strands of DNA. Nature. 1987 May 14;327(6118):169–170. doi: 10.1038/327169a0. [DOI] [PubMed] [Google Scholar]
  41. Yandell D. W., Dryja T. P., Little J. B. Somatic mutations at a heterozygous autosomal locus in human cells occur more frequently by allele loss than by intragenic structural alterations. Somat Cell Mol Genet. 1986 May;12(3):255–263. doi: 10.1007/BF01570784. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES