Effects of DNA Double-Strand and Single-Strand Breaks on Intrachromosomal Recombination Events in Cell-Cycle-Arrested Yeast Cells

Corresponding author: Robert H. Schiestl, Department of Molecular and Cellular Toxicology, Harvard School of Public Health, 665 Huntington Ave., Boston, MA 02115, schiestl{at}mbcrr.harvard.edu (E-mail).

Communicating editor: S. JINKS-ROBERTSON

Intrachromosomal recombination between repeated elements can result in deletion (DEL recombination) events. We investigated the inducibility of such intrachromosomal recombination events at different stages of the cell cycle and the nature of the primary DNA lesions capable of initiating these events. Two genetic systems were constructed in Saccharomyces cerevisiae that select for DEL recombination events between duplicated alleles of CDC28 and TUB2. We determined effects of double-strand breaks (DSBs) and single-strand breaks (SSBs) between the duplicated alleles on DEL recombination when induced in dividing cells or cells arrested in G1 or G2. Site-specific DSBs and SSBs were produced by overexpression of the I-Sce I endonuclease and the gene II protein (gIIp), respectively. I-Sce I-induced DSBs caused an increase in DEL recombination frequencies in both dividing and cell-cycle-arrested cells, indicating that G1- and G2-arrested cells are capable of completing DSB repair. In contrast, gIIp-induced SSBs caused an increase in DEL recombination frequency only in dividing cells. To further examine these phenomena we used both -irradiation, inducing DSBs as its most relevant lesion, and UV, inducing other forms of DNA damage. UV irradiation did not increase DEL recombination frequencies in G1 or G2, whereas -rays increased DEL recombination frequencies in both phases. Both forms of radiation, however, induced DEL recombination in dividing cells. The results suggest that DSBs but not SSBs induce DEL recombination, probably via the single-strand annealing pathway. Further, DSBs in dividing cells may result from the replication of a UV or SSB-damaged template. Alternatively, UV induced events may occur by replication slippage after DNA polymerase pausing in front of the damage.

This article has been cited by other articles:

				W. W. Ku, J. Aubrecht, R. J. Mauthe, R. H. Schiestl, and A. J. Fornace Jr Genetic Toxicity Assessment: Employing the Best Science for Human Safety Evaluation Part VII: Why Not Start with a Single Test: A Transformational Alternative to Genotoxicity Hazard and Risk Assessment Toxicol. Sci., September 1, 2007; 99(1): 20 - 25. [Abstract] [Full Text] [PDF]

				A. Herman-Antosiewicz, S. D. Stan, E.-R. Hahm, D. Xiao, and S. V. Singh Activation of a novel ataxia-telangiectasia mutated and Rad3 related/checkpoint kinase 1-dependent prometaphase checkpoint in cancer cells by diallyl trisulfide, a promising cancer chemopreventive constituent of processed garlic Mol. Cancer Ther., April 1, 2007; 6(4): 1249 - 1261. [Abstract] [Full Text] [PDF]

				R. Reliene and R. H. Schiestl Glutathione depletion by buthionine sulfoximine induces DNA deletions in mice Carcinogenesis, February 1, 2006; 27(2): 240 - 244. [Abstract] [Full Text] [PDF]

				C. Di Primio, A. Galli, T. Cervelli, M. Zoppe, and G. Rainaldi Potentiation of gene targeting in human cells by expression of Saccharomyces cerevisiae Rad52 Nucleic Acids Res., August 16, 2005; 33(14): 4639 - 4648. [Abstract] [Full Text] [PDF]

				A. Galli, T. Cervelli, and R. H. Schiestl Characterization of the Hyperrecombination Phenotype of the pol3-t Mutation of Saccharomyces cerevisiae Genetics, May 1, 2003; 164(1): 65 - 79. [Abstract] [Full Text] [PDF]

				C. Rodriguez, J. Tompkin, J. Hazel, and P. L. Foster Induction of a DNA Nickase in the Presence of Its Target Site Stimulates Adaptive Mutation in Escherichia coli J. Bacteriol., October 15, 2002; 184(20): 5599 - 5608. [Abstract] [Full Text] [PDF]

				A.J.R. Bishop, B. Kosaras, N. Carls, R.L. Sidman, and R.H. Schiestl Susceptibility of proliferating cells to benzo[a]pyrene-induced homologous recombination in mice Carcinogenesis, April 1, 2001; 22(4): 641 - 649. [Abstract] [Full Text] [PDF]

				J. E. Hulla, J. E. French, and J. K. Dunnick Chromosome 11 allelotypes reflect a mechanism of chemical carcinogenesis in heterozygous p53-deficient mice Carcinogenesis, January 1, 2001; 22(1): 89 - 98. [Abstract] [Full Text] [PDF]

				J. E. French, G. D. Lacks, C. Trempus, J. K. Dunnick, J. Foley, J. Mahler, R. R. Tice, and R. W. Tennant Loss of heterozygosity frequency at the Trp53 locus in p53-deficient (+/-) mouse tumors is carcinogen-and tissue-dependent Carcinogenesis, January 1, 2001; 22(1): 99 - 106. [Abstract] [Full Text] [PDF]

				A. J.R. Bishop and R. H. Schiestl Homologous recombination as a mechanism for genome rearrangements: environmental and genetic effects Hum. Mol. Genet., October 1, 2000; 9(16): 2427 - 2334. [Abstract] [Full Text] [PDF]