Abstract
The human APOBEC3 family of DNA cytosine deaminases serves as a front-line intrinsic immune response to inhibit the replication of diverse retroviruses. APOBEC3F and APOBEC3G are the most potent factors against HIV-1. As a countermeasure, HIV-1 viral infectivity factor (Vif) targets APOBEC3s for proteasomal degradation. Here we report the crystal structure of the Vif-binding domain in APOBEC3F and a novel assay to assess Vif-APOBEC3 binding. Our results point to an amphipathic surface that is conserved in APOBEC3s as critical for Vif susceptibility in APOBEC3F. Electrostatic interactions likely mediate Vif binding. Moreover, structure-guided mutagenesis reveals a straight ssDNA-binding groove distinct from the Vif-binding site, and an 'aromatic switch' is proposed to explain DNA substrate specificities across the APOBEC3 family. This study opens new lines of inquiry that will further our understanding of APOBEC3-mediated retroviral restriction and provides an accurate template for structure-guided development of inhibitors targeting the APOBEC3-Vif axis.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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APOBEC Deaminases
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Amino Acid Sequence
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Binding Sites
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Crystallography, X-Ray
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Cytidine Deaminase
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Cytosine Deaminase / chemistry*
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Cytosine Deaminase / genetics
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Cytosine Deaminase / metabolism
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DNA, Single-Stranded / chemistry*
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DNA, Single-Stranded / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gene Expression
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HIV-1 / chemistry*
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HIV-1 / metabolism
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Host-Pathogen Interactions
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Humans
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Models, Molecular
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Protein Binding
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Protein Interaction Domains and Motifs
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Static Electricity
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vif Gene Products, Human Immunodeficiency Virus / chemistry*
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vif Gene Products, Human Immunodeficiency Virus / genetics
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vif Gene Products, Human Immunodeficiency Virus / metabolism
Substances
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DNA, Single-Stranded
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Protein Isoforms
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Recombinant Proteins
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vif Gene Products, Human Immunodeficiency Virus
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vif protein, Human immunodeficiency virus 1
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Cytosine Deaminase
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APOBEC Deaminases
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APOBEC3 proteins, human
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Cytidine Deaminase