Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene.
The DNA double-strand break (DSB) is a critical type of damage, and can be induced by both endogenous sources (e.g. errors of oxidative metabolism, transposable elements, programmed meiotic breaks, or perturbation of the DNA replication fork) and exogenous sources (e.g. ionizing radiation or radiomi...
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doaj-f3eecf1a66f249ab97bafca31b82516a2020-11-24T22:57:11ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2015-05-01610.3389/fpls.2015.00357140543Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene.Tomoyuki eFurukawa0Tomoyuki eFurukawa1Karel J Angelis2Anne Bagg Britt3University of California, DavisMikado Kyowa Seed Co. Ltd.Institute of Experimental Botany AS CRUniversity of California, DavisThe DNA double-strand break (DSB) is a critical type of damage, and can be induced by both endogenous sources (e.g. errors of oxidative metabolism, transposable elements, programmed meiotic breaks, or perturbation of the DNA replication fork) and exogenous sources (e.g. ionizing radiation or radiomimetic chemicals). Although higher plants, like mammals, are thought to preferentially repair DSBs via nonhomologous end joining (NHEJ), much remains unclear about plant DSB repair pathways. Our reverse genetic approach suggests that DNA polymerase λ is involved in DSB repair in Arabidopsis. The Arabidopsis T-DNA insertion mutant (atpolλ-1) displayed sensitivity to both gamma-irradiation and treatment with radiomimetic reagents, but not to other DNA damaging treatments. The atpolλ-1 mutant showed a moderate sensitivity to DSBs, while Arabidopsis Ku70 and DNA ligase 4 mutants (atku70-3 and atlig4-2), both of which play critical roles in NHEJ, exhibited a hypersensitivity to these treatments. The atpolλ-1/atlig4-2 double mutant exhibited a higher sensitivity to DSBs than each single mutant, but the atku70/atpolλ-1 showed similar sensitivity to the atku70-3 mutant. We showed that transcription of the DNA ligase 1, DNA ligase 6, and Wee1 genes was quickly induced by BLM in several NHEJ deficient mutants in contrast to wild-type. Finally, the T-DNA transformation efficiency dropped in NHEJ deficient mutants and the lowest transformation efficiency was scored in the atpolλ-1/atlig4-2 double mutant. These results imply that AtPolλ is involved in both DSB repair and DNA damage response pathway.http://journal.frontiersin.org/Journal/10.3389/fpls.2015.00357/fullDNA RepairDNA damage responseDNA polymeraseDouble Strand BreaksNon homologous end joining |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tomoyuki eFurukawa Tomoyuki eFurukawa Karel J Angelis Anne Bagg Britt |
spellingShingle |
Tomoyuki eFurukawa Tomoyuki eFurukawa Karel J Angelis Anne Bagg Britt Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene. Frontiers in Plant Science DNA Repair DNA damage response DNA polymerase Double Strand Breaks Non homologous end joining |
author_facet |
Tomoyuki eFurukawa Tomoyuki eFurukawa Karel J Angelis Anne Bagg Britt |
author_sort |
Tomoyuki eFurukawa |
title |
Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene. |
title_short |
Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene. |
title_full |
Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene. |
title_fullStr |
Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene. |
title_full_unstemmed |
Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration ofa transgene. |
title_sort |
arabidopsis dna polymerase lambda mutant is mildly sensitive to dna double strand breaks but defective in integration ofa transgene. |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Plant Science |
issn |
1664-462X |
publishDate |
2015-05-01 |
description |
The DNA double-strand break (DSB) is a critical type of damage, and can be induced by both endogenous sources (e.g. errors of oxidative metabolism, transposable elements, programmed meiotic breaks, or perturbation of the DNA replication fork) and exogenous sources (e.g. ionizing radiation or radiomimetic chemicals). Although higher plants, like mammals, are thought to preferentially repair DSBs via nonhomologous end joining (NHEJ), much remains unclear about plant DSB repair pathways. Our reverse genetic approach suggests that DNA polymerase λ is involved in DSB repair in Arabidopsis. The Arabidopsis T-DNA insertion mutant (atpolλ-1) displayed sensitivity to both gamma-irradiation and treatment with radiomimetic reagents, but not to other DNA damaging treatments. The atpolλ-1 mutant showed a moderate sensitivity to DSBs, while Arabidopsis Ku70 and DNA ligase 4 mutants (atku70-3 and atlig4-2), both of which play critical roles in NHEJ, exhibited a hypersensitivity to these treatments. The atpolλ-1/atlig4-2 double mutant exhibited a higher sensitivity to DSBs than each single mutant, but the atku70/atpolλ-1 showed similar sensitivity to the atku70-3 mutant. We showed that transcription of the DNA ligase 1, DNA ligase 6, and Wee1 genes was quickly induced by BLM in several NHEJ deficient mutants in contrast to wild-type. Finally, the T-DNA transformation efficiency dropped in NHEJ deficient mutants and the lowest transformation efficiency was scored in the atpolλ-1/atlig4-2 double mutant. These results imply that AtPolλ is involved in both DSB repair and DNA damage response pathway. |
topic |
DNA Repair DNA damage response DNA polymerase Double Strand Breaks Non homologous end joining |
url |
http://journal.frontiersin.org/Journal/10.3389/fpls.2015.00357/full |
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