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Loop-extrusion-mediated plasmid DNA cleavage by the bacterial SMC Wadjet complex.

Biswajit Pradhan1, Amar Deep2, Jessica König1

  • 1Max Planck Institute of Biophysics, 60438 Frankfurt am Main, Germany.

Molecular Cell
|December 3, 2024
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Summary

Structural maintenance of chromosomes (SMC) complexes like Wadjet use DNA loop extrusion to recognize and cleave plasmid DNA, defending against transformation. This mechanism is conserved across life.

Keywords:
DNA loop extrusionSMC complexesWadjetbacterial defense systemplasmid restrictionsingle-molecule imaging

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Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • Structural maintenance of chromosomes (SMC) complexes are crucial for genome organization and maintenance in all life forms.
  • Prokaryotic Wadjet complexes, similar to MukBEF, prevent plasmid transformation by cleaving plasmid DNA, but the mechanism is unknown.

Purpose of the Study:

  • To elucidate the molecular mechanism of plasmid recognition and cleavage by the Wadjet complex.
  • To visualize the DNA processing activities of Wadjet using single-molecule imaging.

Main Methods:

  • In vitro single-molecule imaging techniques were employed.
  • Surface-anchored plasmid DNAs were used to observe Wadjet's interaction with DNA.

Main Results:

  • Wadjet functions as a symmetric DNA loop extruder, requiring a dimeric JetABC supercomplex.
  • Wadjet extrudes the entire length of a 44-kb plasmid, stalls, and subsequently cleaves the DNA.
  • This loop extrusion activity is essential for specific plasmid recognition and elimination.

Conclusions:

  • DNA loop extrusion is the mechanism by which Wadjet specifically recognizes and eliminates plasmids.
  • Loop extrusion is an evolutionarily conserved mechanism among SMC complexes across all kingdoms of life.