Abstract
To study the effect of nucleotide excision repair on the spectrum of mutations induced in diploid human fibroblasts by UV light (wavelength, 254 nm), we synchronized repair-proficient cells and irradiated them when the HPRT gene was about to be replicated (early S phase) so that there would be no time for repair in that gene before replication, or in G1 phase 6 h prior to S, and determined the kinds and location of mutations in that gene. As a control, we also compared the spectra of mutations induced in synchronized populations of xeroderma pigmentosum cells (XP12BE cells, which are unable to excise UV-induced DNA damage). Among the 84 mutants sequenced, base substitutions predominated. Of the XP mutants from S or G1 and the repair-proficient mutants from S, approximately 62% were G.C----A.T. In the repair-proficient mutants from G1, 47% were. In mutants from the repair-proficient cells irradiated in S, 71% (10 of 14) of the premutagenic lesions were located in the transcribed strand; with mutants from such cells irradiated in G1, only 20% (3 of 15) were. In contrast, there was no statistically significant difference in the fraction of premutagenic lesions located in the transcribed strand of the XP12BE cells; approximately 75% (24 of 32) of the premutagenic lesions were located in that strand, i.e., 15 of 19 (79%) in the S-phase cells and 9 of 13 (69%) in the G1-phase cells. The switch in strand bias supports preferential nucleotide excision repair of UV-induced damage in the transcribed strand of the HPRT gene.
Full text
PDF







Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bohr V. A., Okumoto D. S., Hanawalt P. C. Survival of UV-irradiated mammalian cells correlates with efficient DNA repair in an essential gene. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3830–3833. doi: 10.1073/pnas.83.11.3830. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bourre F., Renault G., Sarasin A. Sequence effect on alkali-sensitive sites in UV-irradiated SV40 DNA. Nucleic Acids Res. 1987 Nov 11;15(21):8861–8875. doi: 10.1093/nar/15.21.8861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen R. H., Maher V. M., McCormick J. J. Effect of excision repair by diploid human fibroblasts on the kinds and locations of mutations induced by (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene in the coding region of the HPRT gene. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8680–8684. doi: 10.1073/pnas.87.21.8680. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drobetsky E. A., Grosovsky A. J., Glickman B. W. The specificity of UV-induced mutations at an endogenous locus in mammalian cells. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9103–9107. doi: 10.1073/pnas.84.24.9103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glickman B. W., Schaaper R. M., Haseltine W. A., Dunn R. L., Brash D. E. The C-C (6-4) UV photoproduct is mutagenic in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6945–6949. doi: 10.1073/pnas.83.18.6945. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grossmann A., Maher V. M., McCormick J. J. The frequency of mutants in human fibroblasts UV-irradiated at various times during S-phase suggests that genes for thioguanine- and diphtheria toxin-resistance are replicated early. Mutat Res. 1985 Oct;152(1):67–76. doi: 10.1016/0027-5107(85)90047-8. [DOI] [PubMed] [Google Scholar]
- 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]
- Konze-Thomas B., Hazard R. M., Maher V. M., McCormick J. J. Extent of excision repair before DNA synthesis determines the mutagenic but not the lethal effect of UV radiation. Mutat Res. 1982 Jun;94(2):421–434. doi: 10.1016/0027-5107(82)90305-0. [DOI] [PubMed] [Google Scholar]
- Kunz B. A., Pierce M. K., Mis J. R., Giroux C. N. DNA sequence analysis of the mutational specificity of u.v. light in the SUP4-o gene of yeast. Mutagenesis. 1987 Nov;2(6):445–453. doi: 10.1093/mutage/2.6.445. [DOI] [PubMed] [Google Scholar]
- Mellon I., Bohr V. A., Smith C. A., Hanawalt P. C. Preferential DNA repair of an active gene in human cells. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8878–8882. doi: 10.1073/pnas.83.23.8878. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mellon I., Hanawalt P. C. Induction of the Escherichia coli lactose operon selectively increases repair of its transcribed DNA strand. Nature. 1989 Nov 2;342(6245):95–98. doi: 10.1038/342095a0. [DOI] [PubMed] [Google Scholar]
- Mellon I., Spivak G., Hanawalt P. C. Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene. Cell. 1987 Oct 23;51(2):241–249. doi: 10.1016/0092-8674(87)90151-6. [DOI] [PubMed] [Google Scholar]
- Mitchell D. L., Haipek C. A., Clarkson J. M. (6-4)Photoproducts are removed from the DNA of UV-irradiated mammalian cells more efficiently than cyclobutane pyrimidine dimers. Mutat Res. 1985 Jul;143(3):109–112. doi: 10.1016/s0165-7992(85)80018-x. [DOI] [PubMed] [Google Scholar]
- Nelson K. K., Green M. R. Mechanism for cryptic splice site activation during pre-mRNA splicing. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6253–6257. doi: 10.1073/pnas.87.16.6253. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Patton J. D., Rowan L. A., Mendrala A. L., Howell J. N., Maher V. M., McCormick J. J. Xeroderma pigmentosum fibroblasts including cells from XP variants are abnormally sensitive to the mutagenic and cytotoxic action of broad spectrum simulated sunlight. Photochem Photobiol. 1984 Jan;39(1):37–42. doi: 10.1111/j.1751-1097.1984.tb03401.x. [DOI] [PubMed] [Google Scholar]
- Petinga R. A., Andrews A. D., Tarone R. E., Robbins J. H. Typical xeroderma pigmentosum complementation group A fibroblasts have detectable ultraviolet light-induced unscheduled DNA synthesis. Biochim Biophys Acta. 1977 Dec 14;479(4):400–410. doi: 10.1016/0005-2787(77)90033-8. [DOI] [PubMed] [Google Scholar]
- Prelich G., Stillman B. Coordinated leading and lagging strand synthesis during SV40 DNA replication in vitro requires PCNA. Cell. 1988 Apr 8;53(1):117–126. doi: 10.1016/0092-8674(88)90493-x. [DOI] [PubMed] [Google Scholar]
- Ryan P. A., Maher V. M., McCormick J. J. Modification of MCDB 110 medium to support prolonged growth and consistent high cloning efficiency of diploid human fibroblasts. Exp Cell Res. 1987 Oct;172(2):318–328. doi: 10.1016/0014-4827(87)90390-9. [DOI] [PubMed] [Google Scholar]
- Schaaper R. M., Dunn R. L., Glickman B. W. Mechanisms of ultraviolet-induced mutation. Mutational spectra in the Escherichia coli lacI gene for a wild-type and an excision-repair-deficient strain. J Mol Biol. 1987 Nov 20;198(2):187–202. doi: 10.1016/0022-2836(87)90305-6. [DOI] [PubMed] [Google Scholar]
- Sykes R. C., Lin D., Hwang S. J., Framson P. E., Chinault A. C. Yeast ARS function and nuclear matrix association coincide in a short sequence from the human HPRT locus. Mol Gen Genet. 1988 May;212(2):301–309. doi: 10.1007/BF00334700. [DOI] [PubMed] [Google Scholar]
- Tsutsui T., Barrett J. C., Ts'o P. O. Induction of 6-thioguanine- and ouabain-resistant mutations in synchronized Syrian hamster cell cultures during different periods of the S phase. Mutat Res. 1978 Nov;52(2):255–264. doi: 10.1016/0027-5107(78)90146-x. [DOI] [PubMed] [Google Scholar]
- Urlaub G., Mitchell P. J., Ciudad C. J., Chasin L. A. Nonsense mutations in the dihydrofolate reductase gene affect RNA processing. Mol Cell Biol. 1989 Jul;9(7):2868–2880. doi: 10.1128/mcb.9.7.2868. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vrieling H., Van Rooijen M. L., Groen N. A., Zdzienicka M. Z., Simons J. W., Lohman P. H., van Zeeland A. A. DNA strand specificity for UV-induced mutations in mammalian cells. Mol Cell Biol. 1989 Mar;9(3):1277–1283. doi: 10.1128/mcb.9.3.1277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watanabe M., Maher V. M., McCormick J. J. Excision repair of UV- or benzo[a]pyrene diol epoxide-induced lesions in xeroderma pigmentosum variant cells is 'error free'. Mutat Res. 1985 Nov;146(3):285–294. doi: 10.1016/0167-8817(85)90070-7. [DOI] [PubMed] [Google Scholar]
- Yang J. L., Chen R. H., Maher V. M., McCormick J. J. Kinds and location of mutations induced by (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene in the coding region of the hypoxanthine (guanine) phosphoribosyltransferase gene in diploid human fibroblasts. Carcinogenesis. 1991 Jan;12(1):71–75. doi: 10.1093/carcin/12.1.71. [DOI] [PubMed] [Google Scholar]
- Yang J. L., Maher V. M., McCormick J. J. Amplification and direct nucleotide sequencing of cDNA from the lysate of low numbers of diploid human cells. Gene. 1989 Nov 30;83(2):347–354. doi: 10.1016/0378-1119(89)90121-2. [DOI] [PubMed] [Google Scholar]