RecA stimulates sister chromatid exchange and the fidelity of double-strand break repair, but not gene targeting, in plants transformed by Agrobacterium.
Expression of the bacterial RecA protein in plants stimulates homologous recombination in tobacco. Here we show that RecA plays a direct role in DNA strand exchange in vivo. The number of sister chromatid exchanges (SCEs) was increased 2.4-fold over wild type in transgenic tobacco plants expressing a nuclear-targeted RecA (nt-RecA) protein and could not be increased further by DNA damage, which caused a doubling of the baseline SCE frequency in wild-type plants. Although gene targeting requires homologous recombination, the number of targeted gene replacements was not increased markedly by the presence of nt-RecA by using Agrobacterium-mediated transformation. However, the number of double-strand breaks that were repaired at both sides by homologous recombination was increased 3.3-fold. Stimulation of SCE and fidelity of double-strand break repair by nt-RecA, but not by gene targeting, suggests that the stimulatory activity of RecA is linked to active DNA synthesis. Therefore, nascent replication-associated single strands may be a prerequisite for RecA action in plant cells.
Reiss
B
B
Max-Planck-Institut für Züchtungsforschung, Carl-von-Linne-Weg 10, D-50829 Köln, Germany. reiss@mpiz-koeln.mpg.de
Schubert
I
I
Köpchen
K
K
Wendeler
E
E
Schell
J
J
Puchta
H
H
D001483:Base Sequence; D017931:DNA Primers; D004260:DNA Repair; D018390:Gene Targeting; D010944:Plants / Q000235:genetics*; D011938:Rec A Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic; D012231:Rhizobium / Q000502:physiology*; D012854:Sister Chromatid Exchange; D014170:Transformation, Genetic
D017931:DNA Primers; D011938:Rec A Recombinases