Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells
Ultraviolet (UV)-B irradiation leads to DNA damage, cell cycle arrest, growth inhibition, and cell death. To evaluate the UV-B stress–induced changes in plant cells, we developed a model system based on tobacco Bright Yellow-2 (BY-2) cells. Both low-dose UV-B (low UV-B: 740 J m−2) and high-dose UV-B...
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doaj-653a37817f054d8d8d7ba375cfdf79a42020-11-24T22:34:14ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2015-04-01610.3389/fpls.2015.00254135509Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cellsShinya eTakahashi0Shinya eTakahashi1Shinya eTakahashi2Kei H. Kojo3Kei H. Kojo4Natsumaro eKutsuna5Natsumaro eKutsuna6Masaki eEndo7Seiichi eToki8Hiroko eIsoda9Hiroko eIsoda10Seiichiro eHasezawa11University of TsukubaThe University of TokyoUniversity of TsukubaThe University of TokyoLPixelThe University of TokyoLPixelNational Institute of Agrobiological SciencesNational Institute of Agrobiological SciencesUniversity of TsukubaUniversity of TsukubaThe University of TokyoUltraviolet (UV)-B irradiation leads to DNA damage, cell cycle arrest, growth inhibition, and cell death. To evaluate the UV-B stress–induced changes in plant cells, we developed a model system based on tobacco Bright Yellow-2 (BY-2) cells. Both low-dose UV-B (low UV-B: 740 J m−2) and high-dose UV-B (high UV-B: 2960 J m−2) inhibited cell proliferation and induced cell death; these effects were more pronounced at high UV-B. Flow cytometry showed cell cycle arrest within 1 day after UV-B irradiation; neither low- nor high-UV-B–irradiated cells entered mitosis within 12 h. Cell cycle progression was gradually restored in low-UV-B–irradiated cells but not in high-UV-B–irradiated cells. UV-A irradiation, which activates cyclobutane pyrimidine dimer (CPD) photolyase, reduced inhibition of cell proliferation by low but not high UV-B and suppressed high-UV-B–induced cell death. UV-B induced CPD formation in a dose-dependent manner. The amounts of CPDs decreased gradually within 3 days in low-UV-B–irradiated cells, but remained elevated after 3 days in high-UV-B–irradiated cells. Low UV-B slightly increased the number of DNA single-strand breaks detected by the comet assay at 1 day after irradiation, and then decreased at 2 and 3 days after irradiation. High UV-B increased DNA fragmentation detected by the terminal deoxynucleotidyl transferase dUTP nick end labeling assay 1 and 3 days after irradiation. Caffeine, an inhibitor of ataxia telangiectasia mutated (ATM) and ataxia telangiectasia and Rad3-related (ATR) checkpoint kinases, reduced the rate of cell death in high-UV-B–irradiated cells. Our data suggest that low-UV-B–induced CPDs and/or DNA strand-breaks inhibit DNA replication and proliferation of BY-2 cells, whereas larger contents of high-UV-B–induced CPDs and/or DNA strand-breaks lead to cell death.http://journal.frontiersin.org/Journal/10.3389/fpls.2015.00254/fullCell CycleCell DeathDNA Damagecheckpointultraviolet-BBY-2 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Shinya eTakahashi Shinya eTakahashi Shinya eTakahashi Kei H. Kojo Kei H. Kojo Natsumaro eKutsuna Natsumaro eKutsuna Masaki eEndo Seiichi eToki Hiroko eIsoda Hiroko eIsoda Seiichiro eHasezawa |
spellingShingle |
Shinya eTakahashi Shinya eTakahashi Shinya eTakahashi Kei H. Kojo Kei H. Kojo Natsumaro eKutsuna Natsumaro eKutsuna Masaki eEndo Seiichi eToki Hiroko eIsoda Hiroko eIsoda Seiichiro eHasezawa Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells Frontiers in Plant Science Cell Cycle Cell Death DNA Damage checkpoint ultraviolet-B BY-2 |
author_facet |
Shinya eTakahashi Shinya eTakahashi Shinya eTakahashi Kei H. Kojo Kei H. Kojo Natsumaro eKutsuna Natsumaro eKutsuna Masaki eEndo Seiichi eToki Hiroko eIsoda Hiroko eIsoda Seiichiro eHasezawa |
author_sort |
Shinya eTakahashi |
title |
Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells |
title_short |
Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells |
title_full |
Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells |
title_fullStr |
Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells |
title_full_unstemmed |
Differential responses to high- and low-dose ultraviolet-B stress in tobacco Bright Yellow-2 cells |
title_sort |
differential responses to high- and low-dose ultraviolet-b stress in tobacco bright yellow-2 cells |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Plant Science |
issn |
1664-462X |
publishDate |
2015-04-01 |
description |
Ultraviolet (UV)-B irradiation leads to DNA damage, cell cycle arrest, growth inhibition, and cell death. To evaluate the UV-B stress–induced changes in plant cells, we developed a model system based on tobacco Bright Yellow-2 (BY-2) cells. Both low-dose UV-B (low UV-B: 740 J m−2) and high-dose UV-B (high UV-B: 2960 J m−2) inhibited cell proliferation and induced cell death; these effects were more pronounced at high UV-B. Flow cytometry showed cell cycle arrest within 1 day after UV-B irradiation; neither low- nor high-UV-B–irradiated cells entered mitosis within 12 h. Cell cycle progression was gradually restored in low-UV-B–irradiated cells but not in high-UV-B–irradiated cells. UV-A irradiation, which activates cyclobutane pyrimidine dimer (CPD) photolyase, reduced inhibition of cell proliferation by low but not high UV-B and suppressed high-UV-B–induced cell death. UV-B induced CPD formation in a dose-dependent manner. The amounts of CPDs decreased gradually within 3 days in low-UV-B–irradiated cells, but remained elevated after 3 days in high-UV-B–irradiated cells. Low UV-B slightly increased the number of DNA single-strand breaks detected by the comet assay at 1 day after irradiation, and then decreased at 2 and 3 days after irradiation. High UV-B increased DNA fragmentation detected by the terminal deoxynucleotidyl transferase dUTP nick end labeling assay 1 and 3 days after irradiation. Caffeine, an inhibitor of ataxia telangiectasia mutated (ATM) and ataxia telangiectasia and Rad3-related (ATR) checkpoint kinases, reduced the rate of cell death in high-UV-B–irradiated cells. Our data suggest that low-UV-B–induced CPDs and/or DNA strand-breaks inhibit DNA replication and proliferation of BY-2 cells, whereas larger contents of high-UV-B–induced CPDs and/or DNA strand-breaks lead to cell death. |
topic |
Cell Cycle Cell Death DNA Damage checkpoint ultraviolet-B BY-2 |
url |
http://journal.frontiersin.org/Journal/10.3389/fpls.2015.00254/full |
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