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    Tension induces a base-paired overstretched DNA conformation

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    Type
    Article
    Authors
    Bosaeus, Niklas
    El-Sagheer, Afaf H.
    Brown, Tom
    Smith, Steven B.
    Akerman, Bjorn
    Bustamante, Carlos
    Norden, Bengt
    Date
    2012
    Permanent link to this record
    http://hdl.handle.net/10754/671281
    
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    Abstract
    Mixed-sequence DNA molecules undergo mechanical overstretching by approximately 70% at 60 �pN. Since its initial discovery 15 �y ago, a debate has arisen as to whether the molecule adopts a new form [Cluzel P, et al. (1996) $^{Science} 271:792–794; Smith SB, Cui Y, Bustamante C (1996) $^{Science} 271:795 799], or simply denatures under tension [van Mameren J, et al. (2009) Proc Natl Acad Sci USA 106:18231–18236]. Here, we resolve this controversy by using optical tweezers to extend small 60�bp single DNA duplex molecules whose base content can be designed at will. We show that when AT content is high (70%), a force-induced denaturation of the DNA helix ensues at 62 �pN that is accompanied by an extension of the molecule of approximately 70%. By contrast, GC-rich sequences (60% GC) are found to undergo a reversible overstretching transition into a distinct form that is characterized by a 51% extension and that remains base-paired. For the first time, results proving the existence of a stretched basepaired form of DNA can be presented. The extension observed in the reversible transition coincides with that produced on DNA by binding of bacterial RecA and human Rad51, pointing to its possible relevance in homologous recombination.
    Citation
    Bosaeus, N., El-Sagheer, A. H., Brown, T., Smith, S. B., Åkerman, B., Bustamante, C., & Nordén, B. (2012). Tension induces a base-paired overstretched DNA conformation. Proceedings of the National Academy of Sciences, 109(38), 15179–15184. doi:10.1073/pnas.1213172109
    Sponsors
    This work was funded by grants to B.N. from the European Research Council and King Abdullah University of Science and Technology.
    Publisher
    Proceedings of the National Academy of Sciences
    Journal
    Proceedings of the National Academy of Sciences
    DOI
    10.1073/pnas.1213172109
    Additional Links
    http://www.pnas.org/lookup/doi/10.1073/pnas.1213172109
    ae974a485f413a2113503eed53cd6c53
    10.1073/pnas.1213172109
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