Structural basis of HIV-1 capsid recognition by PF74 and CPSF6

Akash Bhattacharya, Steven L. Alam, Thomas Fricke, Kaneil Zadrozny, Jaroslaw Sedzicki, Alexander B. Taylor, Borries Demeler, Owen Pornillos, Barbie K. Ganser-Pornillos, Felipe Diaz-Griffero, Dmitri N. Ivanov, Mark Yeager

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86 Scopus citations

Abstract

Upon infection of susceptible cells by HIV-1, the conical capsid formed by ∼250 hexamers and 12 pentamers of the CA protein is delivered to the cytoplasm. The capsid shields the RNA genome and proteins required for reverse transcription. In addition, the surface of the capsid mediates numerous host-virus interactions, which either promote infection or enable viral restriction by innate immune responses. In the intact capsid, there is an intermolecular interface between the N-terminal domain (NTD) of one subunit and the C-terminal domain (CTD) of the adjacent subunit within the same hexameric ring. The NTD-CTD interface is critical for capsid assembly, both as an architectural element of the CA hexamer and pentamer and as a mechanistic element for generating lattice curvature. Here we report biochemical experiments showing that PF-3450074 (PF74), a drug that inhibits HIV-1 infection, as well as host proteins cleavage and polyadenylation specific factor 6 (CPSF6) and nucleoporin 153 kDa (NUP153), bind to the CA hexamer with at least 10-fold higher affinities compared with nonassembled CA or isolated CA domains. The crystal structure of PF74 in complex with the CA hexamer reveals that PF74 binds in a preformed pocket encompassing the NTD-CTD interface, suggesting that the principal inhibitory target of PF74 is the assembled capsid. Likewise, CPSF6 binds in the same pocket. Given that the NTD-CTD interface is a specificmolecular signature of assembled hexamers in the capsid, binding of NUP153 at this site suggests that key features of capsid architecture remain intact upon delivery of the preintegration complex to the nucleus.

Original languageEnglish (US)
Pages (from-to)18625-18630
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume111
Issue number52
DOIs
StatePublished - Dec 30 2014

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Keywords

  • Drug discovery
  • Fluorescence polarization
  • HIV-1 CA protein
  • Isothermal calorimetry
  • X-ray crystallography

ASJC Scopus subject areas

  • General

Cite this

Bhattacharya, A., Alam, S. L., Fricke, T., Zadrozny, K., Sedzicki, J., Taylor, A. B., Demeler, B., Pornillos, O., Ganser-Pornillos, B. K., Diaz-Griffero, F., Ivanov, D. N., & Yeager, M. (2014). Structural basis of HIV-1 capsid recognition by PF74 and CPSF6. Proceedings of the National Academy of Sciences of the United States of America, 111(52), 18625-18630. https://doi.org/10.1073/pnas.1419945112