Structural dynamics of individual Holliday junctions.

S. A. McKinney, A. C. Déclais, D. M. Lilley, T Ha (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    288 Citations (Scopus)

    Abstract

    The four-way DNA (Holliday) junction is the central intermediate of genetic recombination, but the dynamic aspects of this important structure are presently unclear. Although transitions between alternative stacking conformers have been predicted, conventional kinetic studies are precluded by the inability to synchronize the junction in a single conformer in bulk solution. Using single-molecule fluorescence methodology we have been able to detect these transitions. The sequence dependence, the influence of counterions and measured energetic barriers indicate that the conformer transition and branch migration processes share the unstacked, open structure as the common intermediate but have different rate-limiting steps. Relative rates indicate that multiple conformer transitions occur at each intermediate step of branch migration, allowing the junction to reach conformational equilibrium. This provides a mechanism whereby the sequence-dependent conformational bias could determine the extent of genetic exchange upon junction resolution.
    Original languageEnglish
    Pages (from-to)94-97
    JournalNature Structural and Molecular Biology
    Volume10
    Issue number2
    DOIs
    Publication statusPublished - Feb 2003

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