Repair of ultraviolet radiation damage in xeroderma pigmentosum cells belonging to complementation group F

doi:10.1016/0027-5107(81)90110-X

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis

Volume 80, Issue 2, February 1981, Pages 381-388

https://doi.org/10.1016/0027-5107(81)90110-X Get rights and content

Abstract

DNA-repair characteristics of xeroderma pigmentosum belonging to complementation group F were investigated. The cells exhibited an intermediate level of repair as measured in terms of (1) disappearance of T4 endonuclease-V-susceptible sites from DNA, (2) formation of ultraviolet-induced strand breaks in DNA, and (3) ultraviolet-induced unscheduled DNA synthesis during post-irradiation incubation. The impaired ability of XP3YO to perform unscheduled DNA synthesis was restored, to half the normal level, by the concomitant treatment with T4 endonuclease V and ultraviolet-inactivated Sendai virus. It is suggested that xeroderma pigmentosum cells of group F may be defective, at least in part, in the incision step of excision repair.

References (15)

S. Arase et al.
A sixth complementation group in xeroderma pigmentosum
Mutation Res.
(1979)
W. Keijzer et al.
A seventh complementation group in excision-deficient xeroderma pigmentosum
Mutation Res.
(1979)
K. Minton et al.
The ultraviolet endonuclease of bacteriophage T4; further characterization
J. Biol. Chem.
(1975)
A.A. van Zeeland
Introduction of T4 endonuclease V into frozen and thawed mammalian cells for the determination of removal of UV induced photoproducts
S. Yasuda et al.
Further purification and characterization of T4 endonuclease V
Biochim. Biophys. Acta
(1976)
B. Zelle et al.
Repair of UV-endonuclease-susceptible sites in the 7 complementation groups of xeroderma pigmentosum A through G
Mutation Res.
(1979)
W.C. Dunn et al.
Inhibition of DNA excision repair in human cells by arabinofuranosyl cytosine; effect on normal and xeroderma pigmentosum cells
Mol. Pharmacol.
(1979)

There are more references available in the full text version of this article.

Cited by (41)

A DNA repair gene of Caenorhabditis elegans: A homolog of human XPF
2004, DNA Repair
The xeroderma pigmentosum complementation group F (XPF) protein is a structure-specific endonuclease in a complex with ERCC1 and is essential for nucleotide excision repair (NER). We report a single cDNA of Caenorhabditis elegans (C. elegans) encoding highly similar protein to human XPF and other XPF members. We propose to name the corresponding C. elegans gene xpf. Messenger RNA for C. elegans xpf is 5′-tagged with a SL2 splice leader, suggesting an operon-like expression for xpf. Using RNAi, we showed that loss of C. elegans xpf function caused hypersensitivity to ultra-violet (UV) irradiation, as observed in enhanced germ cell apoptosis and increased embryonic lethality. This study suggests that C. elegans xpf is conserved in evolution and plays a role in the repair of UV-damaged DNA in C. elegans.
Disruption of mouse ERCC1 results in a novel repair syndrome with growth failure, nuclear abnormalities and senescence
1997, Current Biology
Background: The structure-specific ERCC1/XPF endonuclease complex that contains the ERCC1 and XPF subunits is implicated in the repair of two distinct types of lesions in DNA: nucleotide excision repair (NER) for ultraviolet-induced lesions and bulky chemical adducts; and recombination repair of the very genotoxic interstrand cross-links.
Results: Here, we present a detailed analysis of two types of mice with mutations in ERCC1, one in which the gene is ‘knocked out’, and one in which the encoded protein contains a seven amino-acid carboxy-terminal truncation. In addition to the previously reported symptoms of severe runting, abnormalities of liver nuclei and greatly reduced lifespan (which appeared less severe in the truncation mutant), both types of ERCC1-mutant mouse exhibited an absence of subcutaneous fat, early onset of ferritin deposition in the spleen, kidney malfunction, gross abnormalities of ploidy and cytoplasmic invaginations in nuclei of liver and kidney, and compromised NER and cross-link repair. We also found that heterozygosity for ERCC1 mutations did not appear to provide a selective advantage for chemically induced tumorigenesis. An important clue to the cause of the very severe ERCC1-mutant phenotypes is our finding that ERCC1-mutant cells undergo premature replicative senescence, unlike cells from mice with a defect only in NER.
Conclusions: Our results strongly suggest that the accumulation in ERCC1-mutant mice of endogenously generated DNA interstrand cross-links, which are normally repaired by ERCC1-dependent recombination repair, underlies both the early onset of cell cycle arrest and polyploidy in the liver and kidney. Thus, our work provides an insight into the molecular basis of ageing and highlights the role of ERCC1 and interstrand DNA cross-links.
Detection of Nucleotide Excision Repair Incisions in Human Fibroblasts by Immunostaining for PCNA
1995, Experimental Cell Research
During nucleotide excision repair, damaged DNA is incised on both sides of a lesion and an oligomer containing the damage is excised and replaced by repair DNA synthesis. The latter step is accomplished in vitro by proteins that include the DNA polymerase accessory factor PCNA, which binds to DNA ends to initiate repair synthesis. An increased association of PCNA with nuclei occurs after UV irradiation of nonreplicating DNA in normal human fibroblasts, probably following incision of damaged DNA. This property was used to detect the catalysis of nucleotide excision repair incisions in damaged DNA in vivo, by immunostaining of quiescent human fibroblasts with the widely available PC10 antibody. We summarize here a comprehensive survey of PCNA immunostaining in repair-defective xeroderma pigmentosum (XP) cells in comparison to normal cells. XP-A and XP-G cells were completely defective in staining for PCNA 30 min after UV irradiation. This strongly suggests that XP-A and XP-G proteins are absolutely required in cells before any incisions can be formed in damaged DNA. XP-B, XP-C, XP-D, and XP-F cells showed an intermediate level of staining for PCNA after UV irradiation, indicative of partial incision capacity in those cells. UV-irradiated XP-E and XP-V cells showed normal PCNA immunostaining levels, consistent with evidence that the corresponding factors are not essential for the incision step of repair. The results provide further evidence for the involvement of PCNA in the repair process in vivo and give an alternative to traditional approaches for measurement of nucleotide excision repair capability.
Use of in vivo and in vitro assays for the characterization of mammalian excision repair and isolation of repair proteins
1990, Mutation Research-DNA Repair
Elucidation of the molecular mechanism of mammalian nucleotide excision repair requires the availability of purified proteins, DNA substrates with defined lesions and suitable repair assays. Repair assays introduced in recent years vary from testing individual steps and successions of steps in vitro to systems that closely reflect the entire process in vivo. In the first part of this review, an in vivo microinjection system is discussed. The second part of the article reviews an in vitro system for study of repair synthesis promoted by cell extracts. Both systems can be utilized as assays during the purification of protein factors that complement repair-defective xeroderma pigmentosum cells. The effect of purified repair proteins from other organisms on mammalian repair is also considered.
Microinjection of T4 endonuclease V produced by a synthetic denV gene stimulates unscheduled DNA synthesis in both xeroderma pigmentosum and normal cells
1989, Mutation Research-DNA Repair
A structural gene for T4 endonuclease V was constructed by ligating synthetic oligonucleotides. The endonuclease V was overproduced in E. coli under control of the E. coli tryptophan promoter and purified to apparent homogeneity. The product had comparable DNA glycosylase and apurinic/apyrimidinic (AP) endonuclease activities to the natural enzyme in vitro. When this endonuclease V was microinjected into the cytoplasm of xeroderma pigmentosum (XP) cells of complementation group A, B, C, D, F, G or H, unscheduled DNA synthesis (UDS) above the residual level was detected in all the cells at a dose of about 10³ molecules following UV irradiation. The grain numbers of UDS in these XP cells increased with increase in the dose of enzyme and reached a plateau at the normal cell level on introduction of about 10⁴ molecules. Introduction of more enzyme into either XP cells or normal human cells did not increase the grain number under regular labelling conditions (2.5 h, 37°C). In normal mouse cells, introduction of the enzyme increased the grain number more than 4-fold under the same conditions during at least 8.5 h following UV irradiation. Furthermore, with a labelling time of 30 min, the enzyme more than doubled the grain number even in normal human cells.
Reduced DNA repair synthesis in healthy women having first degree relatives with breast cancer
1987, European Journal of Cancer and Clinical Oncology
The DNA repair synthesis induced by UV-C irradiation was studied in unstimulated lymphocytes of 64 healthy women whose mothers, or sisters, or mothers and sisters had had breast cancer. For comparison we took 48 control women. As the parameter for the determination of DNA repair synthesis the incorporation of [³H]thymidine in the presence of 2 mM hydroxyurea was taken. The levels of [³H]thymidine incorporation were reduced by 19 of the 29 women whose mothers, in 17 of the 25 women whose sisters and in nine of the 10 women whose mothers and sisters had had breast cancer. By comparison a decreased level was found in only seven of the 48 control women. This difference between the controls and women having first degree relatives with breast cancer was significant in each group. In an earlier study a redued DNA repair synthesis in breast cancer patients was established. The present findings suggest that DNA repair synthesis may be one of the factors involved in the genesis of breast cancer.

View all citing articles on Scopus

View full text

Repair of ultraviolet radiation damage in xeroderma pigmentosum cells belonging to complementation group F

Abstract

Mutation Res.

Mutation Res.

J. Biol. Chem.

Biochim. Biophys. Acta

Mutation Res.

Inhibition of DNA excision repair in human cells by arabinofuranosyl cytosine; effect on normal and xeroderma pigmentosum cells

Mol. Pharmacol.