Differential roles of proteins involved in migration of Holliday junctions on recombination and tolerance to DNA damaging agents in Rhizobium etli

The recombination genes involved in Holliday junction migration (ruvB, recG, radA) and heteroduplex editing (mutS) were studied in the α-proteobacterium Rhizobium etli. The genes were interrupted with a loxPSp interposon and R. etli mutants, either single or in combination, were constructed by marker exchange. Our results show that these systems play a differential role in sensitivity to DNA damaging agents and recombination in R. etli. RuvB appears to be the main system for tolerance toward agents instigating single- or double-strand breaks (such as UV light, methyl methanesulphonate and nalidixic acid) while the RecG and RadA systems play minor roles in tolerance to these agents. Using five different recombination assays, we have found that a ruvB null mutant showed a notable reduction in recombination proficiency, while a radA mutant was only weakly affected. A null mutation in recG had the opposite effect, enhancing recombination in most of our assays. This effect was more clearly seen in an assay that measured recombination between divergent sequences (i.e. homeologous), but is unaffected by inactivation of mutS. These data indicate that RecG in R. etli limits intra- and intergenomic plasticity.