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Deficiency of both classical and alternative end‐joining pathways leads to a synergistic defect in double‐strand break repair but not to an increase in homology‐dependent gene targeting in Arabidopsis

Merker, Laura 1; Feller, Laura 1; Dorn, Annika 1; Puchta, Holger 1
1 Joseph Gottlieb Kölreuter Institut für Pflanzenwissenschaften (JKIP), Karlsruher Institut für Technologie (KIT)

Abstract:

In eukaryotes, double-strand breaks (DSBs) are either repaired by homologous recombination (HR) or non-homologous end-joining (NHEJ). In somatic plant cells, HR is very inefficient. Therefore, the vast majority of DSBs are repaired by two different pathways of NHEJ. The classical (cNHEJ) pathway depends on the heterodimer KU70/KU80, while polymerase theta (POLQ) is central to the alternative (aNHEJ) pathway. Surprisingly, Arabidopsis plants are viable, even when both pathways are impaired. However, they exhibit severe growth retardation and reduced fertility. Analysis of mitotic anaphases indicates that the double mutant is characterized by a dramatic increase in chromosome fragmentation due to defective DSB repair. In contrast to the single mutants, the double mutant was found to be highly sensitive to the DSB-inducing genotoxin bleomycin. Thus, both pathways can complement for each other efficiently in DSB repair. We speculated that in the absence of both NHEJ pathways, HR might be enhanced. This would be especially attractive for gene targeting (GT) in which predefined changes are introduced using a homologous template. Unexpectedly, the polq single mutant as well as the double mutant showed significantly lower GT frequencies in comparison to wildtype plants. ... mehr


Verlagsausgabe §
DOI: 10.5445/IR/1000167572
Veröffentlicht am 25.01.2024
Originalveröffentlichung
DOI: 10.1111/tpj.16604
Scopus
Zitationen: 6
Web of Science
Zitationen: 5
Dimensions
Zitationen: 5
Cover der Publikation
Zugehörige Institution(en) am KIT Joseph Gottlieb Kölreuter Institut für Pflanzenwissenschaften (JKIP)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 04.2024
Sprache Englisch
Identifikator ISSN: 0960-7412, 1365-313X
KITopen-ID: 1000167572
Erschienen in The Plant Journal
Verlag John Wiley and Sons
Band 118
Heft 1
Seiten 242–254
Vorab online veröffentlicht am 05.01.2024
Schlagwörter genome engineering/editing, DNA repair and processing, DNA replication, DNA recombination
Nachgewiesen in Web of Science
Dimensions
Scopus
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