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PMID:22860689

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Citation

Roth, N, Klimesch, J, Dukowic-Schulze, S, Pacher, M, Mannuss, A and Puchta, H' (2012) The requirement for recombination factors differs considerably between different pathways of homologous double-strand break repair in somatic plant cells. Plant J. '

Abstract

In recent years, multiple factors involved in DNA double-strand break (DSB) repair have been characterised in Arabidopsis thaliana. Using homologous sequences in somatic cells, DSBs are mainly repaired by two different pathways: synthesis-dependent strand annealing (SDSA) and single-strand annealing (SSA). By applying recombination substrates in which recombination is initiated by the induction of a site-specific DSB by the homing endonuclease I-SceI, we were able to characterise the involvement of different factors in both pathways. The nucleases MRE11 and COM1, both involved in DSB end processing, were not required for either SDSA or SSA in our assay system. Both SDSA and SSA were even more efficient without MRE11, in accordance with the fact that a loss of MRE11 might negatively affect the efficiency of non-homologous end joining. Loss of the classical recombinase RAD51 or its two paralogues RAD51C and XRCC3, as well as the SWI2/SNF2 remodelling factor RAD54, resulted in a drastic deficiency in SDSA but had hardly any influence on SSA, confirming that a strand exchange reaction is only required for SDSA. The helicase FANCM, which is postulated to be involved in the stabilisation of recombination intermediates, is surprisingly not only needed for SDSA but to a lesser extent also for SSA. Both SSA and SDSA were affected only weakly when the SMC6B protein, implicated in sister chromatid recombination, was absent, indicating that SSA and SDSA are in most cases intrachromatid recombination reactions.

Links

PubMed Online version:10.1111/j.1365-313X.2012.05119.x

Keywords


Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ARATH:F4HYE4

acts_upstream_of_or_within

GO:0000724: double-strand break repair via homologous recombination

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:F4IW45

acts_upstream_of_or_within

GO:0045003: double-strand break repair via synthesis-dependent strand annealing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:RAD51

acts_upstream_of_or_within

GO:0045003: double-strand break repair via synthesis-dependent strand annealing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:CHR25

involved_in

GO:0045003: double-strand break repair via synthesis-dependent strand annealing

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:CHR25

acts_upstream_of_or_within

GO:0045003: double-strand break repair via synthesis-dependent strand annealing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:XRCC3

acts_upstream_of_or_within

GO:0045003: double-strand break repair via synthesis-dependent strand annealing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:RAD51

involved_in

GO:0045003: double-strand break repair via synthesis-dependent strand annealing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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