Induction of telomerase activity by in vivo X-irradiation of mouse splenocytes and its possible role in chromosome healing.
Telomeres serve as protective caps for the chromosome ends. They are one of the functional elements required for the stable transmission of eukaryotic chromosomes. Telomerase, a ribonucleoprotein, stabilises the telomere length by adding telomere repeats on to chromosome ends. Telomeres and telomerase can play a role in the formation of chromosome aberrations and especially in healing of the chromosome or chromatid breaks produced by radiation-induced DNA damage. Telomerase-independent processes also appear to be capable of capping broken chromosome ends. We have studied the expression of telomerase, telomere status and chromosome rearrangements in mouse splenocytes following different doses (0.5, 1.0, 2.0 or 3.0 Gy) of X-irradiation in vivo up to 224 days post-exposure. A dose-dependent increase in telomerase activity up to 2 Gy X-ray exposure was observed immediately after irradiation. The increased enzyme activity was detected even up to day 224 post-irradiation, the last time point studied, especially at higher doses (2 Gy and 3 Gy). A significant difference in average telomere length, measured by quantitative fluorescence in situ hybridisation (Q-FISH) on metaphase chromosomes, noticed immediately after irradiation indicates terminal deletion or altered telomere chromatin. However, telomere length was not statistically significant from the control at the later time points studied. Presence of telomere repeats at the chromosomal breakage sites revealed by FISH with peptide nucleic acid (PNA) telomeric probe indicates a possible role of telomerase-dependent or independent processes in chromosome healing and telomere capture in mammalian cells. We found that approximately 25 to 50% of the newly formed telomeres at the breakage sites are in the range of 200 bp to 1 kb, which might suggest that these repeats could have been added by telomerase which showed a corresponding increase following irradiation.
Hande
M P
MP
Department of Radiation Genetics and Chemical Mutagenesis, Leiden University, Leiden, Netherlands. prakash@terryfox.ubc.ca
Lansdorp
P M
PM
Natarajan
A T
AT
D000818:Animals; D002478:Cells, Cultured; D002842:Chromatids / Q000528:radiation effects; D002869:Chromosome Aberrations / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004260:DNA Repair / Q000235:genetics*; D004790:Enzyme Induction / Q000528:radiation effects*; D005260:Female; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D013154:Spleen / Q000201:enzymology*; D019098:Telomerase / Q000378:metabolism*; D016615:Telomere / Q000235:genetics; D014916:Whole-Body Irradiation; D014965:X-Rays
D015342:DNA Probes; D019098:Telomerase