A novel nuclease is the executing part of a bacterial plasmid defense system - Archive ouverte HAL Access content directly
Preprints, Working Papers, ... Year :

A novel nuclease is the executing part of a bacterial plasmid defense system

(1) , (2, 3) , (4) , (1) , (5) , (6) , (4) , (4) , (7, 8)
1
2
3
4
5
6
7
8

Abstract

Cells are continuously facing the risk of taking up foreign DNA that can compromise genomic and cellular integrity. Therefore, bacteria are in a constant arms race with mobile genetic elements such as phages, transposons and plasmids. They have developed several active strategies against invading DNA molecules that can be seen as a bacterial ‘innate immune system’. Anti-phage systems are usually organized in ‘defense islands’ and can consist of restriction-modification (R-M) systems, CRISPR-Cas, and abortive infection (Abi) systems. Despite recent advances in the field, much less is known about plasmid defense systems. We have recently identified the MksBEFG system in Corynebacterium glutamicum as a novel plasmid defense system which comprises homologues of the condensin system MukFEB. Here, we investigated the molecular arrangement of the MksBEFG complex. Importantly, we identified MksG as a novel nuclease that degrades plasmid DNA and is, thus, the executing part of the system. The crystal structure of MksG revealed a dimeric assembly through its DUF2220 C-terminal domains. This domain is homologous to the TOPRIM domain of the topoisomerase II family of enzymes and contains the corresponding divalent ion binding site that is essential for DNA cleavage in topoisomerases, explaining the in vitro nuclease activity of MksG. We further show that the MksBEF subunits exhibit an ATPase cycle similar to MukBEF in vitro and we reason that this reaction cycle, in combination with the nuclease activity provided by MksG, allows for processive degradation of invading plasmids. Super-resolution localization microscopy revealed that the Mks system is spatially regulated via to the polar scaffold protein DivIVA. Introduction of plasmids increases the diffusion rate and alters the localization of MksG, indicating an activation of the system in vivo .
Fichier principal
Vignette du fichier
2022.10.06.511153v1.full.pdf (887.45 Ko) Télécharger le fichier

Dates and versions

pasteur-03914070 , version 1 (27-12-2022)

Licence

Attribution - NonCommercial - NoDerivatives - CC BY 4.0

Identifiers

Cite

Manuela Weiß, Giacomo Giacomelli, Mathilde Ben Assaya, Finja Grundt, Ahmed Haouz, et al.. A novel nuclease is the executing part of a bacterial plasmid defense system. 2022. ⟨pasteur-03914070⟩

Collections

PASTEUR CNRS
0 View
0 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More