Redox controls RecA protein activity via reversible oxidation of its methionine residues
Cite this dataset
Ezraty, Benjamin et al. (2021). Redox controls RecA protein activity via reversible oxidation of its methionine residues [Dataset]. Dryad. https://doi.org/10.5061/dryad.zpc866t78
Reactive oxygen species (ROS) cause damage to DNA and proteins. The RecA protein plays a central role in the bacterial response to DNA damage. Here we report that RecA itself is also targeted by ROS. In vivo consequences of RecA oxidation include defects in SOS induction, DNA repair efficiency, and P1 transduction. In vitro, oxidized RecA fails to display ATPase activity, DNA strand exchange capacity and formation of nucleofilaments. Consistently, mass spectrometry analysis of oxidized RecA revealed that four out of 9 Met residues have been converted to methionine sulfoxide (Met-O). Specifically, mimicking the oxidation of Met35 by changing it for Gln caused complete loss of function whereas mimicking oxidation of Met164 resulted in constitutive SOS activation and loss of recombination activity. The harmful effects of oxidation of RecA were mitigated by repairing activities of methionine sulfoxide reductases MsrA and MsrB, which suppressed all ROS-induced modifications of RecA activity. This work identifies RecA as a substrate of the anti-ROS activity of the MsrA/B enzymes. These findings indicate that under oxidative stress, MsrA/B maintains a level of reduced functional RecA necessary to carry out both efficient recombination and SOS regulation. The implication of this work is that ROS might inflict damages to DNA both directly and indirectly via hampering RecA repair activity.