title: Mechanism of telomere shortening by oxidative stress.
abstract: We investigated whether oxidative stress, which contributes to aging, accelerates the telomere shortening in human cultured cells. The terminal restriction fragment (TRF) from WI-38 fibroblasts irradiated with UVA (365-nm light) decreased with increasing of the irradiation dose. Furthermore, UVA irradiation dose-dependently increased the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in both WI-38 fibroblasts and HL-60 cells. In order to clarify the mechanism of the acceleration of telomere shortening, we investigated site-specific DNA damage induced by UVA irradiation in the presence of endogenous photosensitizers using (32)P 5' end-labeled DNA fragments containing telomeric oligonucleotide (TTAGGG)(4). UVA irradiation with riboflavin induced 8-oxodG formation in the DNA fragments containing telomeric sequence, and Fpg protein treatment led to chain cleavages at the central guanine of 5'-GGG-3' in telomere sequence. Human 8-oxodG-DNA glycosylase introduces a chain break in a double-stranded oligonucleotide specifically at an 8-oxodG residue. The amount of 8-oxodG formation in DNA fragment containing telomere sequence [5'-CGC(TTAGGG)(7)CGC-3'] was approximately five times more than that in the DNA fragment containing nontelomere sequence [5'-CGC(TGTGAG)(7)CGC-3']. Furthermore, H(2)O(2) plus Cu(II) caused DNA damage, including 8-oxodG formation, specifically at the GGG sequence in the telomere sequence (5'-TTAGGG-3'). It is concluded that the formation of 8-oxodG at the GGG triplet in telomere sequence induced by oxidative stress could participate in acceleration of telomere shortening.
authors.affiliation: Department of Environmental and Molecular Medicine, Mie University School of Medicine, Mie 514-8507, Japan. kawanisi@doc.medic.mie-u.ac.jp
authors.forename: Shosuke
authors.initials: S
authors.lastname: Kawanishi
authors.forename: Shinji
authors.initials: S
authors.lastname: Oikawa
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000375:Aging; D003300:Copper / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D005609:Free Radicals; D018922:HL-60 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009841:Oligonucleotides / Q000737:chemistry; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D012256:Riboflavin / Q000494:pharmacology; D016615:Telomere / Q000648:ultrastructure*; D014466:Ultraviolet Rays
chemical_list: D005609:Free Radicals; D009841:Oligonucleotides; D003300:Copper; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D006861:Hydrogen Peroxide; D003849:Deoxyguanosine; D010100:Oxygen; D012256:Riboflavin