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Using Magnetic Tweezers to Unravel the Mechanism of the G-quadruplex Binding and Unwinding Activities of DHX36 Helicase

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RNA Remodeling Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2209))

Abstract

Single-molecule manipulation methods are useful techniques to probe the interactions of proteins and nucleic acid structures. Here, we describe the magnetic tweezers-based single-molecule investigation of the binding of helicases to G-quadruplex structures and their ATP-dependent unwinding activity, using DHX36 (also known as RHAU and G4R1) helicase and a DNA G-quadruplex structure for an example. We specifically emphasize on the principle and method to probe the interactions between DHX36 and the DNA G-quadruplex in different intermediate states during an ATPase cycle of DHX36, based on detecting the DHX36-induced changes in the lifetime of the DNA G-quadruplex under tension. The principle of the measurement can be broadly extended to the studies of other DNA or RNA G-quadruplex helicases.

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Acknowledgments

This work was supported by the Ministry of Education Academic Research Fund Tier 1 (to J. Y.); The National Research Foundation, Prime Minister’s Office, Singapore and the Ministry of Education under the Research Centres of Excellence programme (to J. Y.); The National Natural Science Foundation of China (21708009 to H.Y.), and the Fundamental Research Fund for the Central Universities (2017KFYXJJ153 to H.Y.).

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Authors and Affiliations

  1. School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Huijuan You

  2. Mechanobiology Institute, National University of Singapore, Singapore, Singapore

    Yu Zhou & Jie Yan

Authors
  1. Huijuan You
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  2. Yu Zhou
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  3. Jie Yan
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Corresponding author

Correspondence to Jie Yan .

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Editors and Affiliations

  1. Centre de Biophysique Moléculaire, CNRS UPR4301, Orleans Cedex 2, France

    Marc Boudvillain

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You, H., Zhou, Y., Yan, J. (2021). Using Magnetic Tweezers to Unravel the Mechanism of the G-quadruplex Binding and Unwinding Activities of DHX36 Helicase. In: Boudvillain, M. (eds) RNA Remodeling Proteins. Methods in Molecular Biology, vol 2209. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0935-4_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0935-4_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0934-7

  • Online ISBN: 978-1-0716-0935-4

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