Home LiteratureArticle Details
PMID: 10409751 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Interstrand cross-links induce DNA synthesis in damaged and undamaged plasmids in mammalian cell extracts.

Molecular and cellular biology ·Vol. 19 ·No. 8 ·1999-08-00 ·Pages 5619-30

Li L, Peterson CA, Lu X, Wei P, Legerski RJ

Abstract

Mammalian cell extracts have been shown to carry out damage-specific DNA repair synthesis induced by a variety of lesions, including those created by UV and cisplatin. Here, we show that a single psoralen interstrand cross-link induces DNA synthesis in both the damaged plasmid and a second homologous unmodified plasmid coincubated in the extract. The presence of the second plasmid strongly stimulates repair synthesis in the cross-linked plasmid. Heterologous DNAs also stimulate repair synthesis to variable extents. Psoralen monoadducts and double-strand breaks do not induce repair synthesis in the unmodified plasmid, indicating that such incorporation is specific to interstrand cross-links. This induced repair synthesis is consistent with previous evidence indicating a recombinational mode of repair for interstrand cross-links. DNA synthesis is compromised in extracts from mutants (deficient in ERCC1, XPF, XRCC2, and XRCC3) which are all sensitive to DNA cross-linking agents but is normal in extracts from mutants (XP-A, XP-C, and XP-G) which are much less sensitive. Extracts from Fanconi anemia cells exhibit an intermediate to wild-type level of activity dependent upon the complementation group. The DNA synthesis deficit in ERCC1- and XPF-deficient extracts is restored by addition of purified ERCC1-XPF heterodimer. This system provides a biochemical assay for investigating mechanisms of interstrand cross-link repair and should also facilitate the identification and functional characterization of cellular proteins involved in repair of these lesions.

MeSH Terms
Animals Cell Line Cell-Free System Cross-Linking Reagents/pharmacology DNA Damage DNA Repair DNA, Recombinant/biosynthesis,drug effects DNA-Binding Proteins/genetics,physiology Endonucleases Fanconi Anemia/genetics,pathology Ficusin/pharmacology HeLa Cells Humans Mammals/genetics,metabolism Mice Plasmids/drug effects,genetics Proteins/genetics,physiology Rad51 Recombinase Recombination, Genetic Tissue Extracts Xeroderma Pigmentosum/genetics,pathology
Chemicals
Cross-Linking Reagents DNA, Recombinant DNA-Binding Proteins Proteins Tissue Extracts xeroderma pigmentosum group F protein RAD51 protein, human Rad51 Recombinase Rad51 protein, mouse ERCC1 protein, human Endonucleases Ercc1 protein, mouse Ficusin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Li L
Departments of Experimental Radiation Oncology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Peterson C A
Lu X
Wei P
Legerski R J
References (51)
51 references, click to expand
  1. Human endonucleolytic incision of DNA 3' and 5' to a site-directed psoralen monoadduct and interstrand cross-link.
    J Biol Chem. 1995 Dec 22;270(51):30709-16 PMID: 8530510
  2. Furocoumarin-photosensitized hydroxylation of guanosine in RNA and DNA.
    Photochem Photobiol. 1995 Apr;61(4):336-40 PMID: 7537887
  3. Removal of psoralen interstrand cross-links from DNA of Escherichia coli: mechanism and genetic control.
    J Mol Biol. 1976 May 5;103(1):39-59 PMID: 785009
  4. A sensitive endonuclease probe for lesions in deoxyribonucleic acid helix structure produced by carcinogenic or mutagenic agents.
    J Biol Chem. 1977 Dec 10;252(23):8740-6 PMID: 925019
  5. Cross-link formation of phage lambda DNA in situ photochemically induced by the furocoumarin derivative angelicin.
    Biochim Biophys Acta. 1980 Apr 30;607(2):215-20 PMID: 6445206
  6. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract.
    Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855-9 PMID: 6933441
  7. Repair of interstrand cross-links in DNA of Saccharomyces cerevisiae requires two systems for DNA repair: the RAD3 system and the RAD51 system.
    Mol Gen Genet. 1981;182(2):196-205 PMID: 7026973
  8. Evidence for DNA double-strand breaks as the critical lesions in yeast cells irradiated with sparsely or densely ionizing radiation under oxic or anoxic conditions.
    Radiat Res. 1981 Dec;88(3):524-32 PMID: 7031753
  9. DNA strand exchange mediated by a RAD51-ssDNA nucleoprotein filament with polarity opposite to that of RecA.
    Cell. 1995 Aug 11;82(3):453-61 PMID: 7634335
  10. Classification of Fanconi anemia patients by complementation analysis: evidence for a fifth genetic subtype.
    Blood. 1995 Sep 15;86(6):2156-60 PMID: 7662964
  11. Cytoplasmic localization of FAC is essential for the correction of a prerepair defect in Fanconi anemia group C cells.
    J Clin Invest. 1996 May 1;97(9):2003-10 PMID: 8621788
  12. Evidence that mammalian cells possess homologous recombinational repair pathways.
    Mutat Res. 1996 Jun 12;363(2):77-88 PMID: 8676928
  13. Double-strand break repair in the absence of RAD51 in yeast: a possible role for break-induced DNA replication.
    Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):7131-6 PMID: 8692957
  14. Molecular analysis of Fanconi anaemia.
    Bioessays. 1996 Jul;18(7):579-85 PMID: 8757936
  15. Nucleotide excision repair genes as determinants of cellular sensitivity to cyclophosphamide analogs.
    Cancer Chemother Pharmacol. 1996;38(5):406-16 PMID: 8765433
  16. Expression cloning of a cDNA for the major Fanconi anaemia gene, FAA.
    Nat Genet. 1996 Nov;14(3):320-3 PMID: 8896563
  17. Positional cloning of the Fanconi anaemia group A gene.
    Nat Genet. 1996 Nov;14(3):324-8 PMID: 8896564
  18. Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro.
    Cell. 1996 Nov 15;87(4):757-66 PMID: 8929543
  19. Beyond DNA cross-linking: history and prospects of DNA-targeted cancer treatment--fifteenth Bruce F. Cain Memorial Award Lecture.
    Cancer Res. 1996 Dec 15;56(24):5533-46 PMID: 8971150
  20. DNA-PK: at the cross-roads of biochemistry and genetics.
    Mutat Res. 1997 Jun 9;384(1):1-14 PMID: 9201268
  21. Evidence for at least eight Fanconi anemia genes.
    Am J Hum Genet. 1997 Oct;61(4):940-4 PMID: 9382107
  22. Initiation of DNA interstrand cross-link repair in humans: the nucleotide excision repair system makes dual incisions 5' to the cross-linked base and removes a 22- to 28-nucleotide-long damage-free strand.
    Mol Cell Biol. 1997 Dec;17(12):6822-30 PMID: 9372913
  23. The Fanconi anaemia proteins, FAA and FAC, interact to form a nuclear complex.
    Nat Genet. 1997 Dec;17(4):487-90 PMID: 9398857
  24. XRCC2 and XRCC3, new human Rad51-family members, promote chromosome stability and protect against DNA cross-links and other damages.
    Mol Cell. 1998 May;1(6):783-93 PMID: 9660962
  25. Chromosomal rearrangements occur in S. cerevisiae rfa1 mutator mutants due to mutagenic lesions processed by double-strand-break repair.
    Mol Cell. 1998 Jul;2(1):9-22 PMID: 9702187
  26. Is Fanconi anemia caused by a defect in the processing of DNA damage?
    Mutat Res. 1998 Aug 7;408(2):75-90 PMID: 9739810
  27. Cleavage at a V(D)J recombination signal requires only RAG1 and RAG2 proteins and occurs in two steps.
    Cell. 1995 Nov 3;83(3):387-95 PMID: 8521468
  28. The fate of 8-methoxypsoralen photoinduced crosslinks in nuclear and mitochondrial yeast DNA: comparison of wild-type and repair-deficient strains.
    Proc Natl Acad Sci U S A. 1982 Mar;79(6):1722-6 PMID: 6281782
  29. Genetic control of excision of Saccharomyces cerevisiae interstrand DNA cross-links induced by psoralen plus near-UV light.
    Mol Cell Biol. 1982 Aug;2(8):939-48 PMID: 6752694
  30. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  31. Molecular cloning of a human DNA repair gene.
    Nature. 1984 Aug 2-8;310(5976):425-9 PMID: 6462228
  32. Simian virus 40 DNA replication in vitro.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):6973-7 PMID: 6095264
  33. Defective DNA cross-link removal in Chinese hamster cell mutants hypersensitive to bifunctional alkylating agents.
    Cancer Res. 1985 Apr;45(4):1737-43 PMID: 3919945
  34. Psoralens as photoactive probes of nucleic acid structure and function: organic chemistry, photochemistry, and biochemistry.
    Annu Rev Biochem. 1985;54:1151-93 PMID: 2411210
  35. Action mechanism of ABC excision nuclease on a DNA substrate containing a psoralen crosslink at a defined position.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8077-81 PMID: 3534882
  36. Complementation of the xeroderma pigmentosum DNA repair defect in cell-free extracts.
    Cell. 1988 Apr 8;53(1):97-106 PMID: 3349527
  37. Use of psoralen-modified oligonucleotides to trap three-stranded RecA-DNA complexes and repair of these cross-linked complexes by ABC excinuclease.
    J Biol Chem. 1988 Oct 15;263(29):15110-7 PMID: 2844801
  38. In vitro repair of psoralen-DNA cross-links by RecA, UvrABC, and the 5'-exonuclease of DNA polymerase I.
    J Biol Chem. 1989 Apr 25;264(12):6755-65 PMID: 2708342
  39. Human nucleotide excision repair in vitro: repair of pyrimidine dimers, psoralen and cisplatin adducts by HeLa cell-free extract.
    Nucleic Acids Res. 1989 Jun 26;17(12):4471-84 PMID: 2748330
  40. The Chinese hamster V79 cell mutant V-H4 is phenotypically like Fanconi anemia cells.
    Somat Cell Mol Genet. 1990 Nov;16(6):575-81 PMID: 2267631
  41. RecA-dependent incision of psoralen-crosslinked DNA by (A)BC excinuclease.
    Nucleic Acids Res. 1991 Feb 11;19(3):657-63 PMID: 2011535
  42. Cloning of cDNAs for Fanconi's anaemia by functional complementation.
    Nature. 1992 Apr 30;356(6372):763-7 PMID: 1574115
  43. Mutant rodent cell lines sensitive to ultraviolet light, ionizing radiation and cross-linking agents: a comprehensive survey of genetic and biochemical characteristics.
    Mutat Res. 1993 Jan;293(2):99-118 PMID: 7678147
  44. Evidence for at least four Fanconi anaemia genes including FACC on chromosome 9.
    Nat Genet. 1992 Jun;1(3):196-8 PMID: 1303234
  45. Fanconi anemia diagnosis and the diepoxybutane (DEB) test.
    Exp Hematol. 1993 Jun;21(6):731-3 PMID: 8500573
  46. A mammalian protein complex that repairs double-strand breaks and deletions by recombination.
    J Biol Chem. 1993 Jul 15;268(20):15070-9 PMID: 8392064
  47. The Fanconi anemia polypeptide FACC is localized to the cytoplasm.
    Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6712-6 PMID: 7517562
  48. Localization of Fanconi anemia C protein to the cytoplasm of mammalian cells.
    Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):7975-9 PMID: 8058745
  49. Hematologic abnormalities in Fanconi anemia: an International Fanconi Anemia Registry study.
    Blood. 1994 Sep 1;84(5):1650-5 PMID: 8068955
  50. RAD1 and RAD10, but not other excision repair genes, are required for double-strand break-induced recombination in Saccharomyces cerevisiae.
    Mol Cell Biol. 1995 Apr;15(4):2245-51 PMID: 7891718
  51. Repair of DNA containing interstrand crosslinks in Escherichia coli: sequential excision and recombination.
    Proc Natl Acad Sci U S A. 1973 Apr;70(4):1064-8 PMID: 4577788
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.8.5619
Published
1999-08-00
Pages
5619-30
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC84414
Subset
IM
Grants
NCI NIH HHS · CA75160 · United States
NCI NIH HHS · R01 CA075160 · United States
NCI NIH HHS · CA52461 · United States
NCI NIH HHS · R01 CA052461 · United States
NCI NIH HHS · R01 CA076172 · United States
NCI NIH HHS · R29 CA076172 · United States
NCI NIH HHS · CA76172 · United States
External Links
PubMed source DOI: 10.1128/MCB.19.8.5619 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2