Small-angle x-ray and neutron scattering of MexR and its complex with DNA supports a conformational selection binding model

In this work, we used small-angle x-ray and neutron scattering to reveal the shape of the protein-DNA complex of the Pseudomonas aeruginosa transcriptional regulator MexR, a member of the multiple antibiotics resistance regulator (MarR) family, when bound to one of its native DNA binding sites. Several MarR-like proteins, including MexR, repress the expression of efflux pump proteins by binding to DNA on regulatory sites overlapping with promoter regions. When expressed, efflux proteins self-assemble to form multiprotein complexes and actively expel highly toxic compounds out of the host organism. The muta-tional pressure on efflux-regulating MarR family proteins is high since deficient DNA binding leads to constitutive expression of efflux pumps and thereby supports acquired multidrug resistance. Understanding the functional outcome of such mutations and their effects on DNA binding has been hampered by the scarcity of structural and dynamic characterization of both free and DNA-bound MarR proteins. Here, we show how combined neutron and x-ray small-angle scattering of both states in solution support a conformational selection model that enhances MexR asymmetry in binding to one of its promoter-overlapping DNA binding sites.

Automated summary

In this study, we used small-angle x-ray and neutron scattering techniques to investigate the shape of the protein-DNA complex formed by the MexR transcriptional regulator in Pseudomonas aeruginosa. MexR belongs to the MarR family of proteins that repress the expression of efflux pump proteins by binding to specific DNA sites. Through our experiments, we demonstrate how the combination of neutron and x-ray scattering supports a conformational selection model for MexR binding to its DNA site.