Shortened telomeres in murine scid cells expressing mutant hRAD54 coincide with reduction in recombination at telomeres.
Murine severe combined immunodeficiency (scid) cells are characterized by defective Prkdc (DNA-PKcs), one of the key genes involved in the repair of DNA double-strand breaks. Interestingly, scid mice are not null mutants and their cells are likely to show low DNA-PKcs activity. Prkdc is also involved in telomere maintenance and in contrast to mice genetically engineered to lack Prkdc (i.e. null mutants), which show complete absence of DNA-PKcs activity, loss of telomere capping function and normal telomere length, cells from scid mice show not only loss of telomere capping function but also abnormally elongated telomeres. Here we demonstrate that telomere elongation observed in murine scid cells can be reversed by expressing mutant hRAD54, a protein involved in homologous recombination. In addition, we measured recombination rates at telomeres using chromosome orientation fluorescence in situ hybridization (CO-FISH) and found that these are elevated in scid cells in comparison with control cells, or significantly reduced in scid cells expressing mutant hRAD54. Similarly, recombination rates at telomeres are reduced in scid cells following introduction of functional Prkdc. Since expression of mutant hRAD54 and restoration of functional Prkdc in scid cells cause the same effects, i.e. telomere shortening and reduced recombination rates at telomeres, these results argue that telomere elongation in scid cells is a complex trait resulting from interactions between homologous recombination mechanisms and DNA-PKcs.
Brunel Institute of Cancer Genetics and Pharmacogenomics, Division of Biosciences, School of Health Sciences and Social Care, Brunel University, Uxbridge, Middlesex UB8 3PH, UK.
Suliman
S
Al-Wahiby
Hiu-Pak
HP
Wong
P
P
Slijepcevic
D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002869:Chromosome Aberrations / Q000187:drug effects; D004265:DNA Helicases; D051747:DNA-Activated Protein Kinase; D004268:DNA-Binding Proteins; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D018345:Mice, Knockout; D016513:Mice, SCID; D016685:Mitomycin / Q000494:pharmacology; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D011995:Recombination, Genetic; D012854:Sister Chromatid Exchange / Q000187:drug effects; D016615:Telomere
D000477:Alkylating Agents; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016685:Mitomycin; D051747:DNA-Activated Protein Kinase; C497264:Prkdc protein, mouse; D004265:DNA Helicases; C103532:RAD54L protein, human