ABSTRACT For naturally competent microorganisms, such as Helicobacter pylori , the steps that permit recombination of exogenous DNA are not fully understood. Immediately downstream of an H. pylori gene ( dprA ) that facilitates high-frequency natural transformation is HP0334 ( dprB ), annotated to be a putative Holliday junction resolvase (HJR). We showed that the HP0334 ( dprB ) gene product facilitates high-frequency natural transformation. We determined the physiologic roles of DprB by genetic analyses. DprB controls in vitro growth, survival after exposure to UV or fluoroquinolones, and intragenomic recombination. dprB ruvC double deletion dramatically decreases both homologous and homeologous transformation and survival after exposure to DNA-damaging agents. Moreover, the DprB protein binds to synthetic Holliday junction structures rather than double-stranded or single-stranded DNA. These results demonstrate that the dprB product plays important roles affecting inter- and intragenomic recombination. We provide evidence that the two putative H. pylori HJRs (DprB and RuvC) have overlapping but distinct functions involving intergenomic (primarily DprB) and intragenomic (primarily RuvC) recombination.