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Journal Articles Life Science Alliance Year : 2022

Binding stoichiometry and structural model of the HIV-1 Rev/Importin β complex

Abstract

ABSTRACT HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein Importin β (Impβ), but how Rev associates with Impβ is poorly understood. Here, we report biochemical, mutational, and biophysical studies of the Impβ/Rev complex. We show that Impβ binds two Rev monomers through independent binding sites, in contrast to the 1:1 binding stoichiometry observed for most Impβ cargos. Peptide scanning data and charge-reversal mutations identify the N-terminal tip of Rev helix α2 within Rev’s arginine-rich motif (ARM) as a primary Impβ-binding epitope. Cross-linking mass spectrometry and compensatory mutagenesis data combined with molecular docking simulations suggest a structural model in which one Rev monomer binds to the C-terminal half of Impβ with Rev helix α2 roughly parallel to the HEAT-repeat superhelical axis, whereas the other monomer binds to the N-terminal half. These findings shed light on the molecular basis of Rev recognition by Impβ and highlight an atypical binding behavior that distinguishes Rev from canonical cellular Impβ cargos.
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Dates and versions

hal-03453423 , version 1 (28-11-2021)
hal-03453423 , version 2 (28-08-2022)

Licence

Attribution - CC BY 4.0

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Didier Spittler, Rose-Laure Indorato, Elisabetta Boeri Erba, Elise Delaforge, Luca Signor, et al.. Binding stoichiometry and structural model of the HIV-1 Rev/Importin β complex. Life Science Alliance, 2022, ⟨10.26508/lsa.202201431⟩. ⟨hal-03453423v2⟩
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