Torsional control of double-stranded DNA branch migration

Xiaoping Yang, Alexander V. Vologodskii, Bing Liu, Börries Kemper, Nadrian C. Seeman

Research output: Contribution to journalArticlepeer-review

Abstract

DNA branched junctions are analogues of Holliday junction recombination intermediates. Partially mobile junctions contain a limited amount of homology flanking the branch point. A partially mobile DNA branched junction has been incorporated into a synthetic double-stranded circular DNA molecule. The junction is flanked by four homologous nucleotide pairs, so that there are five possible locations for the branch point. Two opposite arms of the branched junction are joined to form the circular molecule, which contains 262 nucleotides to the base of the junction. This molecule represents a system whereby torque applied to the circular molecule can have an impact on the junction, by relocating its branch point. Ligation of the molecule produces two topoisomers; about 87% of the product is a relaxed molecule, and the rest is a molecule with one positive supercoil. The position of the branch point is assayed by cleaving the molecule with endonuclease VII. We find that the major site of the branch point in the relaxed topoisomer is at the maximally extruded position in the relaxed molecule. Upon the addition of ethidium, the major site of the branch point migrates to the minimally extruded position.

Original languageEnglish (US)
Pages (from-to)69-83
Number of pages15
JournalBiopolymers
Volume45
Issue number1
DOIs
StatePublished - 1998

Keywords

  • Branch migration
  • Control of DNA structure
  • DNA branched junctions
  • Endonuclease VII
  • Nanomechanical device
  • Superhelical torque

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Biomaterials
  • Organic Chemistry

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