Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

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
. 1986 Nov;83(21):8273–8277. doi: 10.1073/pnas.83.21.8273

Restricted ultraviolet mutational spectrum in a shuttle vector propagated in xeroderma pigmentosum cells.

A Bredberg, K H Kraemer, M M Seidman
PMCID: PMC386910  PMID: 3464953

Abstract

A shuttle vector plasmid, pZ189, carrying a bacterial suppressor tRNA marker gene, was treated with ultraviolet radiation and propagated in cultured skin cells from a patient with the skin-cancer-prone, DNA repair-deficient disease xeroderma pigmentosum and in repair-proficient cells. After replication in the human cells, progeny plasmids were purified. Plasmid survival and mutations inactivating the marker gene were scored by transforming an indicator strain of Escherichia coli carrying a suppressible amber mutation in the beta-galactosidase gene. Plasmid survival in the xeroderma pigmentosum cells was less than that of pZ189 harvested from repair-proficient human cells. The point-mutation frequency in the 150-base-pair tRNA marker gene increased up to 100-fold with ultraviolet dose. Sequence analysis of 150 mutant plasmids revealed that mutations were infrequent at potential thymine-thymine dimer sites. Ninety-three percent of the mutant plasmids from the xeroderma pigmentosum cells showed G X C----A X T transitions, compared to 73% in the normal cells (P less than 0.002). There were significantly fewer transversions (P less than 0.002) (especially G X C----T X A) and multiple base substitutions (P less than 0.00001) than when pZ189 was passaged in repair-proficient cells. The subset of mutational changes that are common to ultraviolet-treated plasmids propagated in both repair-proficient and xeroderma pigmentosum skin cells may be associated with the development of ultraviolet-induced skin cancer in humans.

Full text

PDF
8273

Selected References

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

  1. Bourre F., Sarasin A. Targeted mutagenesis of SV40 DNA induced by UV light. Nature. 1983 Sep 1;305(5929):68–70. doi: 10.1038/305068a0. [DOI] [PubMed] [Google Scholar]
  2. Brash D. E., Haseltine W. A. UV-induced mutation hotspots occur at DNA damage hotspots. Nature. 1982 Jul 8;298(5870):189–192. doi: 10.1038/298189a0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Day R. S., 3rd Studies on repair of adenovirus 2 by human fibroblasts using normal, xeroderma pigmentosum, and xeroderma pigmentosum heterozygous strains. Cancer Res. 1974 Aug;34(8):1965–1970. [PubMed] [Google Scholar]
  5. Foster P. L., Eisenstadt E., Cairns J. Random components in mutagenesis. Nature. 1982 Sep 23;299(5881):365–367. doi: 10.1038/299365a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gearhart P. J., Bogenhagen D. F. Clusters of point mutations are found exclusively around rearranged antibody variable genes. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3439–3443. doi: 10.1073/pnas.80.11.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Glazer P. M., Sarkar S. N., Summers W. C. Detection and analysis of UV-induced mutations in mammalian cell DNA using a lambda phage shuttle vector. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1041–1044. doi: 10.1073/pnas.83.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HOWARD B. D., TESSMAN I. IDENTIFICATION OF THE ALTERED BASES IN MUTATED SINGLE-STRANDED DNA. 3. MUTAGENESIS BY ULTRAVIOLET LIGHT. J Mol Biol. 1964 Aug;9:372–375. doi: 10.1016/s0022-2836(64)80214-x. [DOI] [PubMed] [Google Scholar]
  9. Hauser J., Seidman M. M., Sidur K., Dixon K. Sequence specificity of point mutations induced during passage of a UV-irradiated shuttle vector plasmid in monkey cells. Mol Cell Biol. 1986 Jan;6(1):277–285. doi: 10.1128/mcb.6.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  11. Kraemer K. H., Coon H. G., Petinga R. A., Barrett S. F., Rahe A. E., Robbins J. H. National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA. Proc Natl Acad Sci U S A. 1975 Jan;72(1):59–63. doi: 10.1073/pnas.72.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kraemer K. H., Lee M. M., Scotto J. DNA repair protects against cutaneous and internal neoplasia: evidence from xeroderma pigmentosum. Carcinogenesis. 1984 Apr;5(4):511–514. doi: 10.1093/carcin/5.4.511. [DOI] [PubMed] [Google Scholar]
  13. Kraemer K. H., Slor H. Xeroderma pigmentosum. Clin Dermatol. 1985 Jan-Mar;3(1):33–69. doi: 10.1016/0738-081x(85)90096-3. [DOI] [PubMed] [Google Scholar]
  14. LeClerc J. E., Istock N. L., Saran B. R., Allen R., Jr Sequence analysis of ultraviolet-induced mutations in M13lacZ hybrid phage DNA. J Mol Biol. 1984 Dec 5;180(2):217–237. doi: 10.1016/s0022-2836(84)80001-7. [DOI] [PubMed] [Google Scholar]
  15. Lebkowski J. S., Clancy S., Miller J. H., Calos M. P. The lacI shuttle: rapid analysis of the mutagenic specificity of ultraviolet light in human cells. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8606–8610. doi: 10.1073/pnas.82.24.8606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Lytle C. D., Nikaido O., Hitchins V. M., Jacobson E. D. Host cell reactivation by excision repair is error-free in human cells. Mutat Res. 1982 Jun;94(2):405–412. doi: 10.1016/0027-5107(82)90303-7. [DOI] [PubMed] [Google Scholar]
  18. Maher V. M., Dorney D. J., Mendrala A. L., Konze-Thomas B., McCormick J. J. DNA excision-repair processes in human cells can eliminate the cytotoxic and mutagenic consequences of ultraviolet irradiation. Mutat Res. 1979 Sep;62(2):311–323. doi: 10.1016/0027-5107(79)90087-3. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Peden K. W., Pipas J. M., Pearson-White S., Nathans D. Isolation of mutants of an animal virus in bacteria. Science. 1980 Sep 19;209(4463):1392–1396. doi: 10.1126/science.6251547. [DOI] [PubMed] [Google Scholar]
  21. Protić-Sabljić M., Kraemer K. H. One pyrimidine dimer inactivates expression of a transfected gene in xeroderma pigmentosum cells. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6622–6626. doi: 10.1073/pnas.82.19.6622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Protić-Sabljić M., Whyte D., Fagan J., Howard B. H., Gorman C. M., Padmanabhan R., Kraemer K. H. Quantification of expression of linked cloned genes in a simian virus 40-transformed xeroderma pigmentosum cell line. Mol Cell Biol. 1985 Jul;5(7):1685–1693. doi: 10.1128/mcb.5.7.1685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Sarkar S., Dasgupta U. B., Summers W. C. Error-prone mutagenesis detected in mammalian cells by a shuttle vector containing the supF gene of Escherichia coli. Mol Cell Biol. 1984 Oct;4(10):2227–2230. doi: 10.1128/mcb.4.10.2227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schaaper R. M., Kunkel T. A., Loeb L. A. Infidelity of DNA synthesis associated with bypass of apurinic sites. Proc Natl Acad Sci U S A. 1983 Jan;80(2):487–491. doi: 10.1073/pnas.80.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  29. Strauss B., Rabkin S., Sagher D., Moore P. The role of DNA polymerase in base substitution mutagenesis on non-instructional templates. Biochimie. 1982 Aug-Sep;64(8-9):829–838. doi: 10.1016/s0300-9084(82)80138-7. [DOI] [PubMed] [Google Scholar]
  30. Todd P. A., Glickman B. W. Mutational specificity of UV light in Escherichia coli: indications for a role of DNA secondary structure. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4123–4127. doi: 10.1073/pnas.79.13.4123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wabl M., Burrows P. D., von Gabain A., Steinberg C. Hypermutation at the immunoglobulin heavy chain locus in a pre-B-cell line. Proc Natl Acad Sci U S A. 1985 Jan;82(2):479–482. doi: 10.1073/pnas.82.2.479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weinberg R. A. The action of oncogenes in the cytoplasm and nucleus. Science. 1985 Nov 15;230(4727):770–776. doi: 10.1126/science.2997917. [DOI] [PubMed] [Google Scholar]
  33. Wood R. D., Skopek T. R., Hutchinson F. Changes in DNA base sequence induced by targeted mutagenesis of lambda phage by ultraviolet light. J Mol Biol. 1984 Mar 5;173(3):273–291. doi: 10.1016/0022-2836(84)90121-9. [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