UV-induced DNA repair synthesis in cells of patients with different forms of xeroderma pigmentosum and of heterozygotes

Kleijer, Wim J.; Weerd-Kastelein, E.A. de; Sluyter, M.L.; Keijzer, W.; Wit, Jan de; Bootsma, D.

doi:10.1016/0027-5107(73)90062-6

Cited by 81 publications

(26 citation statements)

References 19 publications

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“…A similar relationship has been found (13,21) for the three groups of De Weerd-Kastelein et al (9,10). In experiments to relate their three complementation groups with our groups, fusion studies in their laboratory (21) and in ours have revealed that only one of their three groups, the one containing their strain XP2RO, is different from all of our four groups.…”

Section: Discussionsupporting

confidence: 87%

“…Ten of the XP strains were from patients of the NIH series (1). These XP strains and strains XPKMISF (11) and XPiLO (12) are identified by the nomenclature for XP strains (13). Fibroblasts were grown without antibiotics in modified Ham's F12 medium (14) supplemented with 5% fetal calf serum at 370 in a 5% C02-95% air incubator with more than 95% humidity.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA.

Kraemer¹,

Coon²,

Petinga³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

150

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Fibroblast strains from 12 patients with xeroderma pigmentosum had lower than normal rates of DNA repair, as determined by autoradiographic studies of ultraviolet-induced unscheduled nuclear DNA synthesis. The nuclei in binuclear cells, obtained by fusing fibroblasts from certain pairs of these strains, had a greater rate of DNA On coverslip cultures not treated with virus, grains over "lightly labeled" (1, 2, 5, 6) nuclei from 100 consecutively observed mononuclear cells were counted. Virus-treated coverslips were scamied under low power for areas containing multinucleate cells that had the most light labeling. In such an area, grains were counted over the 100 nuclei of 50 consecu-59 Abbreviations: XP, xeroderma pigmentosum; UV, ultraviolet.

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Section: Discussionsupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA.

Kraemer¹,

Coon²,

Petinga³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

150

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“…In addition there is a further class of XP patients (termed "XP variants" in this communication) which, while exhibiting the usual clinical symptoms, seem to be completely normal in excision-repair of pyrimidine dimers (9-14). All XP lines examined have normal rates of rejoining of DNA single-strand breaks induced by ionizing radiation (14).Despite very low levels of excision-repair in cells from most XP patients (and also in rodent cell lines), they can nevertheless tolerate the production in their DNA of over 105 pyrimi- In this communication we show that fibroblast cultures from three XP variants have normal levels of excision-repair, but are abnormal in postreplication repair. After UV-irradiation, the time taken for the newly synthesized DNA to attain a high molecular weight similar to that in unirradiated controls is much longer than in normal cells.…”

mentioning

confidence: 80%

Xeroderma pigmentosum cells with normal levels of excision repair have a defect in DNA synthesis after UV-irradiation.

Lehmann¹,

Kirk-Bell²,

Arlett³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

523

241

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Cells cultured from most patients suffering from the sunlight-sensitive hereditary disorder xeroderma pigmentosum are defective in the ability to excise ultraviolet light (UV)-induced pyrimidine dimers from their DNA. There is, however,-one class of these patients whose cells are completely normal in this excision repair process. We have found that these cells have an abnormality in the manner in which DNA is synthesized after UVirradiation. The time taken to convert initially lowmolecular-weight DNA synthesized in UV-irradiated cells into high-molecular-weight DNA similar in size to that in untreated cells is much greater in these variants than in normal cells. Furthermore, this slow conversion of low to high-molecular-weight newly synthesized DNA is drastically inhibited by caffeine, which has no effect in normal cells. Recently, complementation studies have shown that there are several genetically different forms of XP, all deficient in excision-repair (3)(4)(5)(6)(7)(8). In addition there is a further class of XP patients (termed "XP variants" in this communication) which, while exhibiting the usual clinical symptoms, seem to be completely normal in excision-repair of pyrimidine dimers (9-14). All XP lines examined have normal rates of rejoining of DNA single-strand breaks induced by ionizing radiation (14).Despite very low levels of excision-repair in cells from most XP patients (and also in rodent cell lines), they can nevertheless tolerate the production in their DNA of over 105 pyrimi- In this communication we show that fibroblast cultures from three XP variants have normal levels of excision-repair, but are abnormal in postreplication repair. After UV-irradiation, the time taken for the newly synthesized DNA to attain a high molecular weight similar to that in unirradiated controls is much longer than in normal cells. Furthermore, this conversion of low-to high-molecular weight DNA is drastically inhibited by caffeine, which has very little effect in normal human cells. MATERIALS AND METHODSCell Lines used in these experiments were primary fibroblasts from healthy donors and XP patients listed in Table 1.Excision of Pyrimidine Dimers Measured by Loss of UVEndonuclease-Susceptible Sites. This procedure has been described in detail (24). Briefly; fibroblast cells cultured as described in ref. 24 were labeled with [3H]thymidine, UVirradiated, and incubated in the absence of radioactive label for various times. They were then mixed with an equal volume of unirradiated cells whose DNA had been labeled with [14C]-dT. DNA was extracted from the mixed cell population and incubated with or without the UV-specific endonuclease from Micrococcus luteus, which'specifically nicks DNA near pyrimidine dimers (24,25). The DNA products resulting from enzymic attack were centrifuged through 5-20% alkaline sucrose gradients at 40,000 rpm for 135 min at 210 in an SW56 rotor. After centrifugation, fractions were collected, radioactivity was determined, and the weight-average molecular weight of the DNA distributions ...

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“…Both the X P cell strains showed a very low level of unscheduled DNA synthesis. Comparison of the U V stimulated 3H-thymidine incorporation in the X P strains with the incorporation found in the control strain following a dose of 100 erg/mm2, which is not a rate-limiting dose for the control (KLEIJER et al 1973), indicates a residual activity of only 5 in both strains.…”

Section: Unscheduled Dna Synthesismentioning

confidence: 83%

Normal chiasma formation in a male with xeroderma pigmentosum

et al. 2009

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A normal chiasma count was found in first spermatocytes from one patient with the classic symptoms of the genetic disease xeroderma pigmentosum (XP). Skin fibroblasts from this patient and from a brother affected with the same disease demonstrated a decreased ability to perform D N A repair synthesis, after ultraviolet light exposure. These findings indicate that the defective enzyme in XP, probably an incision enzyme, is not involved in the process of crossing-over during meiosis. Maj Hultt!n, Department of Clinical Genetics, Karolinska Hospital, S-10401 Stockholm 60, SwedenIn models explaining the recombination events during the meiotic prophase it is postulated that breaks are introduced into the DNA strands at late zygotene. The subsequent repair involving DNA synthesis and strand rejoining could then lead to non-sister chromatid DNA exchanges. (HOTTA and STERN 1971), wheat and rye (FAVELL and WALKER 1973) by density centrifugation techniques. An endonucleolytic activity and a ligase activity were observed in Lilium in an interval during meiotic prophase restricted to crossing-over (HOWELL and STERN 1971). The endonuclease performs an incision step resulting in single-strand breaks in the DNA, whereas the ligase rejoins nucleotides with specific termini.In some microorganisms and in mammalian cells a DNA repair mechanism, called dark repair, is known to operate after ultraviolet light (UV) irradiation. The steps in this dark repair mechanism do include an incision-, a DNA synthesis-and a rejoining step, resulting in the replacement of the DNA containing the UV induced lesions, presumably pyrimidine dimers. This mechanism was first recognized in bacteria (BOYCE and HOWARD-FLANDERS 1964;SETLOW and CARRIER 1964) but is also operative in higher organisms and man (PAINTER 1970

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UV-induced DNA repair synthesis in cells of patients with different forms of xeroderma pigmentosum and of heterozygotes

Cited by 81 publications

References 19 publications

National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA.

National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA.

Xeroderma pigmentosum cells with normal levels of excision repair have a defect in DNA synthesis after UV-irradiation.

Normal chiasma formation in a male with xeroderma pigmentosum

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