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@article{1210516,
author = {Vu, Giang T.H. and Cao, Hieu X. and Watanabe, Koichi and Hensel, Goetze and Blattner, Frank R. and Kumlehn, Jochen and Schubert, Ingo},
article_location = {Rockville (USA)},
article_number = {5},
doi = {http://dx.doi.org/10.1105/tpc.114.126607},
keywords = {SOMATIC PLANT-CELLS; HOMOLOGOUS RECOMBINATION; PHYSCOMITRELLA-PATENS; GENOME EVOLUTION; SEQUENCES; EXCHANGES; MECHANISM; REPLICATION; CONVERSION; CHOICE},
language = {eng},
issn = {1040-4651},
journal = {The Plant Cell},
title = {Repair of Site-Specific DNA Double-Strand Breaks in Barley Occurs via Diverse Pathways Primarily Involving the Sister Chromatid},
url = {http://www.plantcell.org/content/26/5/2156},
volume = {26},
year = {2014}
}
TY - JOUR
ID - 1210516
AU - Vu, Giang T.H. - Cao, Hieu X. - Watanabe, Koichi - Hensel, Goetze - Blattner, Frank R. - Kumlehn, Jochen - Schubert, Ingo
PY - 2014
TI - Repair of Site-Specific DNA Double-Strand Breaks in Barley Occurs via Diverse Pathways Primarily Involving the Sister Chromatid
JF - The Plant Cell
VL - 26
IS - 5
SP - 2156-2167
EP - 2156-2167
PB - American Society of Plant Physiologists
SN - 10404651
KW - SOMATIC PLANT-CELLS
KW - HOMOLOGOUS RECOMBINATION
KW - PHYSCOMITRELLA-PATENS
KW - GENOME EVOLUTION
KW - SEQUENCES
KW - EXCHANGES
KW - MECHANISM
KW - REPLICATION
KW - CONVERSION
KW - CHOICE
UR - http://www.plantcell.org/content/26/5/2156
L2 - http://www.plantcell.org/content/26/5/2156
N2 - DNA double-strand break (DSB) repair mechanisms differ in their requirements for a homologous repair template and in the accuracy of the result. We aimed to quantify the outcome of repair of a single targeted DSB in somatic cells of young barley (Hordeum vulgare) plants. Amplicon sequencing of three reporter constructs revealed 47 to 58% of reads as repaired via nonhomologous end-joining (NHEJ) with deletions and/or small (1 to 3 bp) insertions. Alternative NHEJ revealed 2 to 5 bp microhomology (15.7% of cases) or new replication-mediated short duplications at sealed breaks. Although deletions outweigh insertions in barley, this bias was less pronounced and deleted sequences were shorter than in Arabidopsis thaliana. Between 17 and 33% of reads likely represent restoration of the original sequence. Depending on the construct, 20 to 33% of reads arose via gene conversion (homologous recombination). Remarkably, < 1 to > 8% of reads apparently display synthesis-dependent strand annealing linked with NHEJ, inserting 4 to 61 bp, mostly originating from the surrounding of breakpoints. Positional coincidence of > 81% of sister chromatid exchanges with target loci is unprecedented for higher eukaryotes and indicates that most repair events for staggered DSBs, at least in barley, involve the sister chromatid and occur during S or G2 phase of the cell cycle.
ER -
VU, Giang T.H., Hieu X. CAO, Koichi WATANABE, Goetze HENSEL, Frank R. BLATTNER, Jochen KUMLEHN and Ingo SCHUBERT. Repair of Site-Specific DNA Double-Strand Breaks in Barley Occurs via Diverse Pathways Primarily Involving the Sister Chromatid. \textit{The Plant Cell}. Rockville (USA): American Society of Plant Physiologists, 2014, vol.~26, No~5, p.~2156-2167. ISSN~1040-4651. Available from: https://dx.doi.org/10.1105/tpc.114.126607.
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