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Ring-shaped replicative helicase encircles double-stranded DNA during unwinding.

Sihwa Joo1,2, Bong H Chung1,2,3, Mina Lee4

  • 1BioNanoTechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea.

Nucleic Acids Research
|October 31, 2019
PubMed
Summary

Bovine papillomavirus E1 helicase unwinds DNA poorly, with tension hindering its action. Researchers observed E1 encircling double-stranded DNA, revealing new insights into its unwinding mechanism.

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Area of Science:

  • Molecular biology
  • Biophysics
  • Virology

Background:

  • Ring-shaped replicative helicases are essential for DNA replication.
  • The unwinding mechanism of these helicases is not fully understood.
  • Bovine papillomavirus E1 is a well-studied model replicative helicase.

Purpose of the Study:

  • To investigate the DNA unwinding mechanism of the bovine papillomavirus E1 helicase at the single-molecule level.
  • To elucidate the role of DNA tension and strand interaction in E1 helicase activity.

Main Methods:

  • Single-molecule biophysics using magnetic tweezers.
  • Characterization of E1 helicase unwinding kinetics and force dependence.
  • Comparison of wild-type E1 with a DNA-binding domain-deficient construct.

Main Results:

  • E1 functions as a poorly processive helicase with a low unwinding rate.
  • DNA tension impedes E1 unwinding, suggesting strong interaction with both DNA strands.
  • E1 was observed to encircle double-stranded DNA at high forces (26-30 pN).

Conclusions:

  • The study reveals E1's limited processivity and sensitivity to DNA tension.
  • The discovery of E1 encircling dsDNA provides novel mechanistic insights.
  • Two potential modes of E1 encircling during unwinding are proposed.