Hypersensitivity of mouse NEIL1-knockdown cells to hydrogen peroxide during S phase.
Oxidative base damage occurs spontaneously due to reactive oxygen species generated as byproducts of respiration and other pathological processes in mammalian cells. Many oxidized bases are mutagenic and/or toxic, and most are repaired through the base excision repair pathway. Human endonuclease VIII-like protein 1 (hNEIL1) is thought to play an important role during the S phase of the cell cycle by removing oxidized bases in DNA replication fork-like (bubble) structures, and the protein level of hNEIL1 is increased in S phase. Compared with hNEIL1, there is relatively little information on the properties of the mouse ortholog mNEIL1. Since mouse cell nuclei lack endonuclease III-like protein (NTH) activity, in contrast to human cell nuclei, mNEIL1 is a major DNA glycosylase for repair of oxidized pyrimidines in mouse nuclei. In this study, we made mNEIL1-knockdown cells using an shRNA expression vector and examined the cell cycle-related variation in hydrogen peroxide (H2O2) sensitivity. Hypersensitivity to H2O2 caused by mNEIL1 knockdown was more significant in S phase than in G1 phase, suggesting that mNEIL1 has an important role during S phase, similarly to hNEIL1.
Department of Advanced Pathobiology, Graduate School of Life & Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 598-8531, Japan yamamoto@vet.osakafu-u.ac.jp.
Ryohei
R
Yamamoto
Department of Advanced Pathobiology, Graduate School of Life & Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 598-8531, Japan.
Yukari
Y
Ohshiro
Department of Advanced Pathobiology, Graduate School of Life & Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 598-8531, Japan.
Tatsuhiko
T
Shimotani
Department of Advanced Pathobiology, Graduate School of Life & Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 598-8531, Japan.
Mizuki
M
Yamamoto
Department of Advanced Pathobiology, Graduate School of Life & Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 598-8531, Japan.
Satoshi
S
Matsuyama
Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan.
Hiroshi
H
Ide
Department of Advanced Pathobiology, Graduate School of Life & Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ourai Kita, Izumisano, Osaka 598-8531, Japan.
Kihei
K
Kubo
D000818:Animals; D002478:Cells, Cultured; D004249:DNA Damage; D045647:DNA Glycosylases / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects; D016193:G1 Phase / Q000187:drug effects; D055785:Gene Knockdown Techniques; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007739:L Cells; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics; D016196:S Phase / Q000187:drug effects*
D034741:RNA, Small Interfering; D006861:Hydrogen Peroxide; D045647:DNA Glycosylases; C473634:NEIL1 protein, human; C481347:Neil1 protein, mouse
BER; NEIL1; cell cycle; glycosylase; mouse