Exploration of DNA processing features unravels novel properties of ICE conjugation in Gram-positive bacteria
2022
Laroussi, Haifa | Aoudache, Yanis | Robert, Emilie | Libante, Virginie | Thiriet, Louise | Mias-Lucquin, Dominique | Douzi, Badreddine | Roussel, Yvonne | Chauvot de Beauchene, Isaure | Soler, Nicolas | Leblond-Bourget, Nathalie | Dynamique des Génomes et Adaptation Microbienne (DynAMic) ; Université de Lorraine (UL)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Computational Algorithms for Protein Structures and Interactions (CAPSID) ; Centre Inria de l'Université de Lorraine ; Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria)-Department of Complex Systems, Artificial Intelligence & Robotics (LORIA - AIS) ; Laboratoire Lorrain de Recherche en Informatique et ses Applications (LORIA) ; Institut National de Recherche en Informatique et en Automatique (Inria)-CentraleSupélec-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche en Informatique et en Automatique (Inria)-CentraleSupélec-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-Laboratoire Lorrain de Recherche en Informatique et ses Applications (LORIA) ; Institut National de Recherche en Informatique et en Automatique (Inria)-CentraleSupélec-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-CentraleSupélec-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS) | Regional Operational Program of the European Regional Development Fund “FEDER-FSE Lorraine et Massif des Vosges 2014-2020” | ANR-15-IDEX-0004,LUE,Isite LUE(2015)
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Show more [+] Less [-]English. Integrative and conjugative elements (ICEs) are important drivers of horizontal gene transfer in prokaryotes. They are responsible for antimicrobial resistance spread, a major current health concern. ICEs are initially processed by relaxases that recognize the binding site of oriT sequence and nick at a conserved nic site. The ICESt3/Tn916/ICEBs1 superfamily, which is widespread among Firmicutes, encodes uncanonical relaxases belonging to a recently identified family called MOB T. This family is related to the rolling circle replication initiators of the Rep trans family. The nic site of these MOB T relaxases is conserved but their DNA binding site is still unknown. Here, we identified the bind site of RelSt3, the MOB T relaxase from ICESt3. Unexpectedly, we found this bind site distantly located from the nic site. We revealed that the binding of the RelSt3 N-terminal HTH domain is required for efficient nicking activity. We also deciphered the role of RelSt3 in the initial and final stages of DNA processing during conjugation. Especially, we demonstrated a strand transfer activity, and the formation of covalent DNA-relaxase intermediate for a MOB T relaxase.
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