DNA-measuring Wadjet SMC ATPases restrict smaller circular plasmids by DNA cleavage

Published: 1 December 2022| Version 1 | DOI: 10.17632/8dyprtxwt8.1
Contributors:
Hon Liu, Florian Roisné-Hamelin, Stephan Gruber

Description

Structural maintenance of chromosomes (SMC) complexes fold DNA by loop extrusion to support chromosome segregation and genome maintenance. Wadjet systems (JetABCD/MksBEFG/EptABCD) are derivative SMC complexes with roles in bacterial immunity against selfish DNA. Here, we show that JetABCD restricts circular plasmids with an upper size limit of about 100 kb, while a linear plasmid evades restriction. Purified JetABCD complexes cleave circular DNA molecules regardless of DNA helical topology; cleavage is DNA sequence-nonspecific and depends on the SMC ATPase. A cryo-EM structure reveals a distinct JetABC dimer-of-dimers geometry with the two SMC dimers facing in opposite direction—rather than the same as observed with MukBEF. We hypothesize that JetABCD is a DNA-shape-specific endonuclease and propose the ‘total extrusion model’ for DNA cleavage exclusively when extrusion of an entire plasmid has been completed by a JetABCD complex. Total extrusion cannot be achieved on the larger chromosome, explaining how self-DNA may evade processing.

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Institutions

  • Universite de Lausanne

Categories

Gel, Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, Agarose Gel Electrophoresis

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