Structure and dynamics of supercoil-stabilized DNA cruciforms

Luda S. Shlyakhtenko, Vladimir N. Potaman, Richard R. Sinden, Yuri L. Lyubchenko

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

Understanding DNA function requires knowledge of the structure of local, sequence-dependent conformations that can be dramatically different from the B-form helix. One alternative DNA conformation is the cruciform, which has been shown to have a critical role in the initiation of DNA replication and the regulation of transcription in certain systems. In addition, cruciforms provide a model system for structural studies of Holliday junctions, intermediates in homologous DNA recombination. Cruciforms are not thermodynamically stable in linear DNA due to branch point migration, which makes their study using many biophysical techniques problematic. Atomic Force Microscopy (AFM) was applied to visualize cruciforms in negatively supercoiled plasmid DNA. Cruciforms are seen as clear-cut extrusions on the DNA filament with the lengths of the arms consistent with the size of the hairpins expected from a 106 bp inverted repeat. The cruciform exists in two different conformations, an extended one with the angle of ca. 180°between the hairpin arms and a compact, X-type conformation, with acute angles between the hairpin arms and the main DNA strands. The ratio of molecules with the different conformations of cruciforms depends on ionic conditions. In the presence of high salt or Mg cations, a compact, X-type conformation is highly preferable. Remarkably, the X-conformation was highly mobile allowing the cruciform arms to adopt a parallel orientation. The structure observed is consistent with a model of the Holliday junction with a parallel orientation of the exchanging strands.

Original languageEnglish (US)
Pages (from-to)61-72
Number of pages12
JournalJournal of Molecular Biology
Volume280
Issue number1
DOIs
StatePublished - Jul 3 1998

Fingerprint

Cruciform DNA
DNA
Nucleic Acid Conformation
Superhelical DNA
Atomic Force Microscopy
Homologous Recombination
Structural Models
DNA Replication
Cations
Plasmids
Salts

Keywords

  • Atomic Force Microscopy
  • Cruciform conformation
  • DNA supercoiling
  • Holliday junction
  • Inverted repeats in DNA

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Structure and dynamics of supercoil-stabilized DNA cruciforms. / Shlyakhtenko, Luda S.; Potaman, Vladimir N.; Sinden, Richard R.; Lyubchenko, Yuri L.

In: Journal of Molecular Biology, Vol. 280, No. 1, 03.07.1998, p. 61-72.

Research output: Contribution to journalArticle

Shlyakhtenko, Luda S. ; Potaman, Vladimir N. ; Sinden, Richard R. ; Lyubchenko, Yuri L. / Structure and dynamics of supercoil-stabilized DNA cruciforms. In: Journal of Molecular Biology. 1998 ; Vol. 280, No. 1. pp. 61-72.
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