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TitleDNA-measuring Wadjet SMC ATPases restrict smaller circular plasmids by DNA cleavage.
Journal, issue, pagesMol Cell, Vol. 82, Issue 24, Page 4727-44740.e6, Year 2022
Publish dateDec 15, 2022
AuthorsHon Wing Liu / Florian Roisné-Hamelin / Bertrand Beckert / Yan Li / Alexander Myasnikov / Stephan Gruber /
PubMed AbstractStructural maintenance of chromosome (SMC) complexes fold DNA by loop extrusion to support chromosome segregation and genome maintenance. Wadjet systems (JetABCD/MksBEFG/EptABCD) are derivative SMC ...Structural maintenance of chromosome (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, whereas a linear plasmid evades restriction. Purified JetABCD complexes cleave circular DNA molecules, regardless of the 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.
External linksMol Cell / PubMed:36525956
MethodsEM (single particle)
Resolution3.0 - 3.52 Å
Structure data

15609

8as8

EMDB-15609, PDB-8as8:
E. coli Wadjet JetABC monomer
Method: EM (single particle) / Resolution: 3.0 Å

16020

8bfn

EMDB-16020, PDB-8bfn:
E. coli Wadjet JetABC dimer of dimers
Method: EM (single particle) / Resolution: 3.52 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

Source
  • escherichia coli (E. coli)
KeywordsDNA BINDING PROTEIN / SMC complexes / bacterial immunity / defense system / DNA loop extrusion / Wadjet / JetABCD / DNA cleavage / plasmid restriction / mksBEFG / mobile genetic elements / horizontal gene transfer / eptABCD / mukBEF

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