Abstract
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide–like 3G (APOBEC3G, referred to here as A3G) is a potent antiretroviral host factor against human immunodeficiency virus type 1 (HIV-1). HIV-1 viral infectivity factor (Vif) counteracts A3G by promoting its degradation via the ubiquitin-proteasome pathway. Recent studies demonstrated that protein kinase A (PKA) phosphorylates activation-induced deaminase (AID), another member of the APOBEC3 family. A3G has two putative PKA phosphorylation residues. Here we show that PKA binds and specifically phosphorylates A3G at Thr32 in vitro and in vivo. This phosphorylation event reduces the binding of A3G to Vif and its subsequent ubiquitination and degradation, and thus promotes A3G antiviral activity. Computer-assisted structural modeling and mutagenesis studies suggest that the interaction between A3G Thr32 and Arg24 is crucial for interaction with Vif. These data imply that PKA-mediated phosphorylation of A3G can regulate the interaction between A3G and Vif.
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Acknowledgements
We thank H. Akari (Tukuba Primate Research Center, National Institute of Biomedical Innovation) for the pNLA1-43Vif plasmid, A. Imura (Graduate School of Medicine, Kyoto University) for the anti-GFP mAb and M. Malim (King's College London School of Medicine) for providing the anti-Vif monoclonal antibody (#319) through the AIDS Research and Reference Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Disease, National Institutes of Health. This study was partly supported by grants-in-aid from the Ministry of Education, Culture, Sports, Science and Technology and from the Ministry of Health, Labour and Welfare, Japan. This study was also partly supported by grants-in-aid from the Naito Foundation, from the Mitsubishi Pharma Research Foundation and from The Shimizu Foundation for the Promotion of Immunology Research.
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K.S. designed research, performed research, contributed vital new reagents, analyzed data and wrote the paper; A.T.-K. designed research, analyzed data, wrote the paper and organized research; M.Y. and H.S. performed the structure modeling and wrote the paper; T.I., M.M., K.I. and T.S. prepared the materials and performed a part of research; T.U. analyzed data, drafted the paper and organized research.
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Shirakawa, K., Takaori-Kondo, A., Yokoyama, M. et al. Phosphorylation of APOBEC3G by protein kinase A regulates its interaction with HIV-1 Vif. Nat Struct Mol Biol 15, 1184–1191 (2008). https://doi.org/10.1038/nsmb.1497
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DOI: https://doi.org/10.1038/nsmb.1497
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