J 2014

Repair of Site-Specific DNA Double-Strand Breaks in Barley Occurs via Diverse Pathways Primarily Involving the Sister Chromatid

VU, Giang T.H., Hieu X. CAO, Koichi WATANABE, Goetze HENSEL, Frank R. BLATTNER et. al.

Basic information

Original name

Repair of Site-Specific DNA Double-Strand Breaks in Barley Occurs via Diverse Pathways Primarily Involving the Sister Chromatid

Authors

VU, Giang T.H. (276 Germany), Hieu X. CAO (276 Germany), Koichi WATANABE (276 Germany), Goetze HENSEL (276 Germany), Frank R. BLATTNER (276 Germany), Jochen KUMLEHN (276 Germany) and Ingo SCHUBERT (276 Germany, guarantor, belonging to the institution)

Edition

The Plant Cell, Rockville (USA), American Society of Plant Physiologists, 2014, 1040-4651

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 9.338

RIV identification code

RIV/00216224:14740/14:00077479

Organization unit

Central European Institute of Technology

UT WoS

000338771700030

Keywords in English

SOMATIC PLANT-CELLS; HOMOLOGOUS RECOMBINATION; PHYSCOMITRELLA-PATENS; GENOME EVOLUTION; SEQUENCES; EXCHANGES; MECHANISM; REPLICATION; CONVERSION; CHOICE

Tags

International impact, Reviewed
Změněno: 26/11/2014 07:46, Martina Prášilová

Abstract

V originále

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.

Links

EE2.3.20.0189, research and development project
Name: Rozvoj výzkumné excelence v oblasti evoluční cytogenomiky, epigenetiky a buněčné signalizace