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MksEF Accessory Proteins Inhibit MksB ATPase Activity and Modulate DNA Substrate Binding.

Pratibha Kumari1,2, Shamila Minnu K C1,2, Teasha Biswas3,2

  • 1Department of Molecular Nutrition, CSIR-Central Food Technological Research Institute (CFTRI), Mysuru, Karnataka 570020, India.

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|September 9, 2025
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Summary
This summary is machine-generated.

This study reveals how the MksBEF complex in Mycobacterium smegmatis organizes chromosomes. Accessory proteins MksE and MksF enhance DNA binding but suppress ATPase activity, offering unique insights into bacterial chromosome dynamics.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Chromosome organization and segregation are vital cellular processes.
  • Mechanisms of bacterial nucleoid organization are not fully understood.
  • Structural Maintenance of Chromosomes (SMC) complexes play key roles in genome management.

Purpose of the Study:

  • Investigate the function of the MksBEF SMC complex in *Mycobacterium smegmatis*.
  • Elucidate the role of MksB, MksE, and MksF in chromosome organization.
  • Understand the regulatory mechanisms of this alternative SMC complex.

Main Methods:

  • Reconstitution of the MksBEF complex.
  • Determination of complex stoichiometry (MksB₂E₄F₂) using gel filtration and ultracentrifugation.
  • DNA-binding assays (gel shift, isothermal titration calorimetry) and fluorescence spectroscopy (ANS) to study protein-DNA interactions and structural changes.

Main Results:

  • MksB binds DNA and is crucial for local chromosome organization.
  • MksE and MksF enhance MksB's DNA-binding affinity through direct interaction.
  • DNA binding induces structural rearrangements in MksB and the MksBEF complex.
  • MksEF suppresses MksB's ATPase activity, a novel regulatory mechanism.

Conclusions:

  • The MksBEF complex represents an alternative SMC system with unique regulatory functions.
  • MksE and MksF modulate MksB activity, impacting DNA binding and ATPase function.
  • Findings advance the understanding of chromosome dynamics and organization in mycobacteria.