1. Structure and function of theLeptospira interrogansperoxide stress regulator (PerR), an atypical PerR devoid of a structural metal-binding site
- Author
-
Patrick Weber, Nadia Benaroudj, Frederick Saul, Sarah Dubrac, Annie Landier, Bénédicte Beaudeau, Raléb Taher, Mounira Kebouchi, Ahmed Haouz, Mathieu Picardeau, Biologie des Spirochètes / Biology of Spirochetes, Institut Pasteur [Paris], Cristallographie (Plateforme) - Crystallography (Platform), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Biologie des Bactéries pathogènes à Gram-positif, This work was supported by the Institut Pasteur and Agence National de la Recherche Grant ANR-08-MIE-018., M. K., R. T., A. L., B. B., and P. W. performed experiments. F. S., A. H., S. D., M. P., and N. B. conceived, performed, and analyzed experiments. N. B. conceived and coordinated the study and wrote the paper. All authors approved the final version of the manuscript.We acknowledge SOLEIL for provision of synchrotron radiation facilities, and we thank the staff of beamline PROXIMA 1 for assistance. We are grateful to Gerald Murray and Ben Adler for providing the perR mutant. We thank Gouzel Karimova, Isabelle Michaud-Soret, and Victor Duarte for critically reading the manuscript and for helpful discussions., ANR-08-MIEN-0018,LEPTOVIR,Les leptospires : de la génétique à la pathogénèse(2008), Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
MESH: Signal Transduction ,0301 basic medicine ,MESH: Leptospira interrogans ,[SDV]Life Sciences [q-bio] ,Mutant ,Regulatory site ,Metal Binding Site ,Cell Cycle Proteins ,spirochaetes ,Biochemistry ,Peroxide ,chemistry.chemical_compound ,Metalloprotein ,oxidative stress ,structural biology ,spirochetes ,MESH: Bacterial Proteins ,Leptospira ,chemistry.chemical_classification ,biology ,Chemistry ,régulation transcriptionnelle ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,transcription regulation ,Leptospira interrogans ,Protein Binding ,Signal Transduction ,Repressor ,Mitosis ,PerR ,reactive oxygen species (ROS) ,Microbiology ,03 medical and health sciences ,MESH: Cell Cycle Proteins ,Bacterial Proteins ,métalloprotéine ,MESH: Protein Binding ,microbiologie ,Molecular Biology ,Binding Sites ,metalloprotein ,stress oxydatif ,Cell Biology ,MESH: Mitosis ,biology.organism_classification ,030104 developmental biology ,MESH: Binding Sites ,Structural biology - Abstract
International audience; Peroxide sensing is essential for bacterial survival during aerobic metabolism and host infection. Peroxide stress regulators (PerRs) are homodimeric transcriptional repressors with each monomer typically containing both structural and regulatory metal-binding sites. PerR binding to gene promoters is controlled by the presence of iron in the regulatory site, and iron-catalyzed oxidation of PerR by H2O2 leads to the dissociation of PerR from DNA. In addition to a regulatory metal, most PerRs require a structural metal for proper dimeric assembly. We present here a structural and functional characterization of the PerR from the pathogenic spirochete Leptospira interrogans, a rare example of PerR lacking a structural metal-binding site. In vivo studies showed that the leptospiral PerR belongs to the peroxide stimulon in pathogenic species and is involved in controlling resistance to peroxide. Moreover, a perR mutant had decreased fitness in other host-related stress conditions, including at 37 °C or in the presence of superoxide anion. In vitro, leptospiral PerR could bind to the perR promoter region in a metal-dependent manner. The crystal structure of the leptospiral PerR revealed an asymmetric homodimer, with one monomer displaying complete regulatory metal coordination in the characteristic caliper-like DNA-binding conformation and the second monomer exhibiting disrupted regulatory metal coordination in an open non-DNA-binding conformation. This structure showed that leptospiral PerR assembles into a dimer in which a metal-induced conformational switch can occur independently in the two monomers. Our study demonstrates that structural metal binding is not compulsory for PerR dimeric assembly and for regulating peroxide stress.
- Published
- 2018
- Full Text
- View/download PDF