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Using synthetic bacterial enhancers to reveal a looping-based mechanism for quenching-like repression

Author

Listed:
  • Michal Brunwasser-Meirom

    (Technion—Israel Institute of Technology)

  • Yaroslav Pollak

    (Russell Berrie Nanotechnology Institute, Technion—Israel Institute of Technology)

  • Sarah Goldberg

    (Technion—Israel Institute of Technology)

  • Lior Levy

    (Technion—Israel Institute of Technology)

  • Orna Atar

    (Technion—Israel Institute of Technology)

  • Roee Amit

    (Technion—Israel Institute of Technology
    Russell Berrie Nanotechnology Institute, Technion—Israel Institute of Technology)

Abstract

We explore a model for ‘quenching-like’ repression by studying synthetic bacterial enhancers, each characterized by a different binding site architecture. To do so, we take a three-pronged approach: first, we compute the probability that a protein-bound dsDNA molecule will loop. Second, we use hundreds of synthetic enhancers to test the model’s predictions in bacteria. Finally, we verify the mechanism bioinformatically in native genomes. Here we show that excluded volume effects generated by DNA-bound proteins can generate substantial quenching. Moreover, the type and extent of the regulatory effect depend strongly on the relative arrangement of the binding sites. The implications of these results are that enhancers should be insensitive to 10–11 bp insertions or deletions (INDELs) and sensitive to 5–6 bp INDELs. We test this prediction on 61 σ54-regulated qrr genes from the Vibrio genus and confirm the tolerance of these enhancers’ sequences to the DNA’s helical repeat.

Suggested Citation

  • Michal Brunwasser-Meirom & Yaroslav Pollak & Sarah Goldberg & Lior Levy & Orna Atar & Roee Amit, 2016. "Using synthetic bacterial enhancers to reveal a looping-based mechanism for quenching-like repression," Nature Communications, Nature, vol. 7(1), pages 1-12, April.
  • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms10407
    DOI: 10.1038/ncomms10407
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