Different pathways of homologous recombination are used for the repair of double-strand breaks within tandemly arranged sequences in the plant genome

@article{citeulike:2801099, abstract = {Summary Different DNA repair pathways that use homologous sequences in close proximity to genomic double-strand breaks (DSBs) result in either an internal deletion or a gene conversion. We determined the efficiency of these pathways in somatic plant cells of transgenic Arabidopsis lines by monitoring the restoration of the beta-glucuronidase (GUS) marker gene. The transgenes contain a recognition site for the restriction endonuclease I-SceI either between direct GUS repeats to detect deletion formation (DGU.US), or within the GUS gene to detect gene conversion using a nearby donor sequence in direct or inverted orientation (DU.GUS and IU.GUS). Without expression of I-SceI, the frequency of homologous recombination (HR) was low and similar for all three constructs. By crossing the different lines with an I-SceI expressing line, DSB repair was induced, and resulted in one to two orders of magnitude higher recombination frequency. The frequencies obtained with the DGU.US construct were about five times higher than those obtained with DU.GUS and IU.GUS, irrespective of the orientation of the donor sequence. Our results indicate that recombination associated with deletions is the most efficient pathway of homologous DSB repair in plants. However, DSB-induced gene conversion seems to be frequent enough to play a significant role in the evolution of tandemly arranged gene families like resistance genes.}, author = {Orel, Nadiya and Kyryk, Anzhela and Puchta, Holger }, citeulike-article-id = {2801099}, doi = {10.1046/j.1365-313X.2003.01832.x}, journal = {The Plant Journal}, keywords = {concerted\_evolution, dsb\_repair, gene\_conversion}, number = {5}, pages = {604--612}, posted-at = {2008-05-15 10:17:40}, priority = {2}, title = {Different pathways of homologous recombination are used for the repair of double-strand breaks within tandemly arranged sequences in the plant genome}, url = {http://www.blackwell-synergy.com/doi/abs/10.1046/j.1365-313X.2003.01832.x}, volume = {35}, year = {2003} }

TY - JOUR ID - citeulike:2801099 L3 - citeulike-article-id:2801099 TI - Different pathways of homologous recombination are used for the repair of double-strand breaks within tandemly arranged sequences in the plant genome JF - The Plant Journal VL - 35 IS - 5 SP - 604 EP - 612 N2 - Summary Different DNA repair pathways that use homologous sequences in close proximity to genomic double-strand breaks (DSBs) result in either an internal deletion or a gene conversion. We determined the efficiency of these pathways in somatic plant cells of transgenic Arabidopsis lines by monitoring the restoration of the beta-glucuronidase (GUS) marker gene. The transgenes contain a recognition site for the restriction endonuclease I-SceI either between direct GUS repeats to detect deletion formation (DGU.US), or within the GUS gene to detect gene conversion using a nearby donor sequence in direct or inverted orientation (DU.GUS and IU.GUS). Without expression of I-SceI, the frequency of homologous recombination (HR) was low and similar for all three constructs. By crossing the different lines with an I-SceI expressing line, DSB repair was induced, and resulted in one to two orders of magnitude higher recombination frequency. The frequencies obtained with the DGU.US construct were about five times higher than those obtained with DU.GUS and IU.GUS, irrespective of the orientation of the donor sequence. Our results indicate that recombination associated with deletions is the most efficient pathway of homologous DSB repair in plants. However, DSB-induced gene conversion seems to be frequent enough to play a significant role in the evolution of tandemly arranged gene families like resistance genes. KW - concerted_evolution KW - dsb_repair KW - gene_conversion AU - Orel, Nadiya AU - Kyryk, Anzhela AU - Puchta, Holger PY - 2003/// UR - http://www.blackwell-synergy.com/doi/abs/10.1046/j.1365-313X.2003.01832.x ER -