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Specific Nucleic Acid Chaperone Activity of HIV-1 Nucleocapsid Protein Deduced from Hairpin Unfolding

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RNA Chaperones

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2106))

Abstract

RNA and DNA hairpin formation and disruption play key regulatory roles in a variety of cellular processes. The 59-nucleotide transactivation response (TAR) RNA hairpin facilitates the production of full-length transcripts of the HIV-1 genome. Yet the stability of this long, irregular hairpin becomes a liability during reverse transcription as 24 base pairs must be disrupted for strand transfer. Retroviral nucleocapsid (NC) proteins serve as nucleic acid chaperones that have been shown to both destabilize the TAR hairpin and facilitate strand annealing with its complementary DNA sequence. Yet it has remained difficult to elucidate the way NC targets and dramatically destabilizes this hairpin while only weakly affecting the annealed product. In this work, we used optical tweezers to measure the stability of TAR and found that adding NC destabilized the hairpin and simultaneously caused a distinct change in both the height and location of the energy barrier. This data was matched to an energy landscape predicted from a simple theory of definite base pair destabilization. Comparisons revealed the specific binding sites found by NC along the irregular TAR hairpin. Furthermore, specific binding explained both the unusual shift in the transition state and the much weaker effect on the annealed product. These experiments illustrate a general method of energy landscape transformation that exposes important physical insights.

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Acknowledgments

This work was supported by grants NIH GM072462 and NSF MCB-1817712 to M.C.W.

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Authors and Affiliations

  1. Department of Physics, Northeastern University, Boston, MA, USA

    Micah J. McCauley & Mark C. Williams

  2. Department of Chemistry and Biochemistry, Ohio State University, Columbus, OH, USA

    Ioulia Rouzina

Authors
  1. Micah J. McCauley
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  2. Ioulia Rouzina
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  3. Mark C. Williams
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Corresponding author

Correspondence to Mark C. Williams .

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Editors and Affiliations

  1. Department of Pediatric Hematology, Oncology and Stem Cell Transplantation, University Hospital of Regensburg, Regensburg, Germany

    Tilman Heise

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McCauley, M.J., Rouzina, I., Williams, M.C. (2020). Specific Nucleic Acid Chaperone Activity of HIV-1 Nucleocapsid Protein Deduced from Hairpin Unfolding. In: Heise, T. (eds) RNA Chaperones. Methods in Molecular Biology, vol 2106. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0231-7_4

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  • DOI: https://doi.org/10.1007/978-1-0716-0231-7_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0230-0

  • Online ISBN: 978-1-0716-0231-7

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