The involvement of key DNA repair pathways in the formation of chromosome rearrangements in embryonic stem cells.
It is vital that embryonic stem (ES) cells, which give rise to the diverse tissues of the mature organism, maintain genetic stability. To understand mechanisms for the prevention and causation of chromosomal instability, we have used spectral karyotyping (SKY) to analyse ES cells from wild-type and repair-gene knockout mice. We chose cells deficient in Ku70 (DNA end joining), Xrcc2 (gene conversion), Ercc1 (single-strand annealing) and Csb (transcription-coupled repair) to represent potentially-important DNA repair pathways, plus an Xpc-deficient line to examine loss of global nucleotide excision repair (NER). Spontaneous and radiation (X-ray or alpha-particle)-induced chromosome changes were assessed to measure the influence of different levels of damage severity on response. We show that most repair pathways (except for global NER) protect against chromosome changes induced by ionizing radiations, while only homology-dependent pathways protect against spontaneous chromosomal change in ES cells. However, for a given level of damage, the prevalence of different types of changes alters in the different repair-deficient lines. Thus, loss of Ercc1, Csb or Ku70 leads to increased fragment formation, but loss of Xrcc2 promotes exchanges between chromosomes. Strikingly, we found that loss of the Csb gene function specifically protects ES cells from complex exchanges, suggesting a role for transcription-associated events in complex exchange formation.
Griffin
Carol
C
Medical Research Council, Radiation & Genome Stability Unit, Harwell, Oxfordshire OX11 0RD, England, UK.
Waard
Harm de
Hd
Deans
Bryan
B
Thacker
John
J
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D043171:Chromosomal Instability; D004249:DNA Damage; D004260:DNA Repair; D004622:Embryo, Mammalian / Q000166:cytology*; D006824:Hybridization, Genetic; D051379:Mice; D018345:Mice, Knockout; D011836:Radiation Tolerance; D011839:Radiation, Ionizing; D012097:Repressor Proteins; D013234:Stem Cells; D014158:Transcription, Genetic
D012097:Repressor Proteins