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Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...
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Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Reversible DNA condensation drives natural transformation.

Joshua I Santiago1, Ishtiyaq Ahmed2, Jeanette Hahn2,3

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Antibiotic resistance spreads via natural transformation. The DNA receptor ComEA uses dynamic oligomers to condense DNA, generating force for periplasmic transport, then decondenses DNA for cytoplasmic entry.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biophysics

Background:

  • Natural transformation is a key mechanism for horizontal gene transfer in bacteria, contributing to the spread of antibiotic resistance.
  • The ComEA protein is a crucial DNA receptor involved in importing extracellular DNA into the bacterial periplasm, but its mechanism remains unclear.

Purpose of the Study:

  • To elucidate the mechanism by which the ComEA DNA receptor facilitates DNA uptake during natural transformation.
  • To investigate the role of ComEA oligomerization and DNA condensation in the DNA transport process.

Main Methods:

  • Single-molecule optical tweezers were employed to measure forces exerted by ComEA on DNA.
  • Electron microscopy was used to visualize ComEA-DNA complexes and their structures.
  • Mutational analysis in Bacillus subtilis assessed the functional importance of ComEA conformations.

Main Results:

  • Geobacillus stearothermophilus ComEA forms dynamic, concentration-dependent oligomers on DNA, switching between bridging and non-bridging conformations.
  • Bridging oligomers, formed at low ComEA concentration, condense DNA and generate sub-piconewton pulling forces.
  • Non-bridging oligomers, formed at high ComEA concentration, decondense DNA and do not generate force.
  • Mutations favoring specific conformations led to transformation deficiency, highlighting the importance of both condensation and decondensation.

Conclusions:

  • ComEA reversibly condenses DNA during natural transformation, utilizing force generation for periplasmic DNA translocation.
  • The transition from force-generating condensation to decondensation by ComEA is essential for subsequent DNA transport into the bacterial cytoplasm.
  • This study reveals a novel mechanism of DNA-protein interaction driving essential bacterial genetic processes.