DNA Replication Is Required To Elicit Cellular Responses to Psoralen-Induced DNA Interstrand Cross-Links

ABSTRACT Following introduction of DNA interstrand cross-links (ICLs), mammalian cells display chromosome breakage or cell cycle delay with a 4N DNA content. To further understand the nature of the delay, previously described as a G2/M arrest, we developed a protocol to generate ICLs during specific intervals of the cell cycle. Synchronous populations of G1, S, and G2 cells were treated with photoactivated 4′-hydroxymethyl-4,5′,8-trimethylpsoralen (HMT) and scored for normal passage into mitosis. In contrast to what was found for ionizing radiation, ICLs introduced during G2 did not result in a G2/M arrest, mitotic arrest, or chromosome breakage. Rather, subsequent passage through S phase was required to trigger both chromosome breakage and arrest in the next cell cycle. Similarly, ICLs introduced during G1 did not cause a G1/S arrest. We conclude that DNA replication is required to elicit the cellular responses of cell cycle arrest and genomic instability after psoralen-induced ICLs. In primary human fibroblasts, the 4N DNA content cell cycle arrest triggered by ICLs was long lasting but reversible. Kinetic analysis suggested that these cells could remove up to ∼2,500 ICLs/genome at an average rate of 11 ICLs/genome/h.

[1]  S. Latt,et al.  Flow cytometric characterization of the response of Fanconi's anemia cells to mitomycin C treatment. , 2005, Cytometry.

[2]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[3]  M. Jasin,et al.  Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations. , 1998, Genes & development.

[4]  P. Jeggo,et al.  Fanconi anemia C gene product plays a role in the fidelity of blunt DNA end-joining. , 1998, Journal of molecular biology.

[5]  S. Vispé,et al.  Overexpression of Rad51 protein stimulates homologous recombination and increases resistance of mammalian cells to ionizing radiation. , 1998, Nucleic acids research.

[6]  A. Sancar,et al.  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 , 1997, Molecular and cellular biology.

[7]  F. Kruyt,et al.  Involvement of the Fanconi's anemia protein FAC in a pathway that signals to the cyclin B/cdc2 kinase. , 1997, Cancer research.

[8]  H. Youssoufian Cytoplasmic localization of FAC is essential for the correction of a prerepair defect in Fanconi anemia group C cells. , 1996, The Journal of clinical investigation.

[9]  D. Averbeck,et al.  Preferential repair in yeast after induction of interstrand cross-links by 8-methoxypsoralen plus UVA. , 1995, Mutation research.

[10]  Liu Vf,et al.  Cell cycle checkpoints and repair of ionizing radiation damage. , 1995 .

[11]  J. Hong,et al.  G2/M-phase arrest and release in ataxia telangiectasia and normal cells after exposure to ionizing radiation. , 1994, Radiation research.

[12]  J. Hamlin Mammalian origins of replication , 1992, BioEssays : news and reviews in molecular, cellular and developmental biology.

[13]  M. Kastan,et al.  Wild-type p53 is a cell cycle checkpoint determinant following irradiation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. H. Vos,et al.  Differential introduction of DNA damage and repair in mammalian genes transcribed by RNA polymerases I and II , 1991, Molecular and cellular biology.

[15]  B. V. Houten Nucleotide excision repair in Escherichia coli. , 1990 .

[16]  H. Crissman,et al.  Synchronization of human diploid fibroblasts at multiple stages of the cell cycle. , 1988, Experimental cell research.

[17]  A. Yeung,et al.  Repair of 4,5',8-trimethylpsoralen monoadducts and cross-links by the Escherichia coli UvrABC endonuclease. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[18]  J. Hearst,et al.  Use of psoralen-modified oligonucleotides to trap three-stranded RecA-DNA complexes and repair of these cross-linked complexes by ABC excinuclease. , 1988, The Journal of biological chemistry.

[19]  J. Mccormick,et al.  Carcinogens can induce homologous recombination between duplicated chromosomal sequences in mouse L cells , 1988, Molecular and cellular biology.

[20]  P. Hanawalt,et al.  Processing of psoralen adducts in an active human gene: Repair and replication of DNA containing monoadducts and interstrand cross-links , 1987, Cell.

[21]  R. Schlegel,et al.  Caffeine-induced uncoupling of mitosis from the completion of DNA replication in mammalian cells. , 1986, Science.

[22]  D. Gruenert,et al.  Repair of psoralen-induced cross-links and monoadducts in normal and repair-deficient human fibroblasts. , 1985, Cancer research.

[23]  P. Rabinovitch,et al.  Endogenous blockage and delay of the chromosome cycle despite normal recruitment and growth phase explain poor proliferation and frequent edomitosis in Fanconi anemia cells. , 1985, American journal of human genetics.

[24]  R. Chanet,et al.  The fate of 8-methoxypsoralen photoinduced crosslinks in nuclear and mitochondrial yeast DNA: comparison of wild-type and repair-deficient strains. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. Resnick,et al.  Molecular recombination and the repair of DNA double-strand breaks in CHO cells. , 1979, Nucleic acids research.

[26]  A. Auerbach,et al.  Susceptibility of Fanconi's anaemia fibroblasts to chromosome damage by carcinogens , 1976, Nature.

[27]  M. Sasaki Is Fanconi's anaemia defective in a process essential to the repair of DNA cross links? , 1975, Nature.

[28]  M. A. Bempong,et al.  Sensitivity of rat testes to inhibitors of nucleic acid synthesis. III. The inheritance of mitomycin C-induced structural rearrangements of chromosomes. , 1975, The Journal of heredity.

[29]  M. A. Bempong,et al.  Sensitivity of rat testis to inhibitors of nucleic acid synthesis. II. Mutagenic action of mitomycin C. , 1973, The Journal of heredity.

[30]  D. Scott,et al.  The induction of chromosome aberrations by nitrogen mustard and its dependence on DNA synthesis , 1969, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[31]  M. Heinrich,et al.  DNA cross-linker-induced G2/M arrest in group C Fanconi anemia lymphoblasts reflects normal checkpoint function. , 1998, Blood.

[32]  V. F. Liu,et al.  Cell cycle checkpoints and repair of ionizing radiation damage. , 1995, Stem cells.

[33]  J. Henriques,et al.  The PSO4 gene is responsible for an error-prone recombinational DNA repair pathway in Saccharomyces cerevisiae. , 1989, Molecular & general genetics : MGG.