@article{1bef52e055724df3a58a97e24217c472,
title = "ARIH2 Is a Vif-Dependent Regulator of CUL5-Mediated APOBEC3G Degradation in HIV Infection",
abstract = "The Cullin-RING E3 ligase (CRL) family is commonly hijacked by pathogens to redirect the host ubiquitin proteasome machinery to specific targets. During HIV infection, CRL5 is hijacked by HIV Vif to target viral restriction factors of the APOBEC3 family for ubiquitination and degradation. Here, using a quantitative proteomics approach, we identify the E3 ligase ARIH2 as a regulator of CRL5-mediated APOBEC3 degradation. The CUL5Vif/CBF{\ss} complex recruits ARIH2 where it acts to transfer ubiquitin directly to the APOBEC3 targets. ARIH2 is essential for CRL5-dependent HIV infectivity in primary CD4+ T cells. Furthermore, we show that ARIH2 cooperates with CRL5 to prime other cellular substrates for polyubiquitination, suggesting this may represent a general mechanism beyond HIV infection and APOBEC3 degradation. Taken together, these data identify ARIH2 as a co-factor in the Vif-hijacked CRL5 complex that contributes to HIV infectivity and demonstrate the operation of the E1-E2-E3/E3-substrate ubiquitination mechanism in a viral infection context. Degradation of APOBEC3 proteins by HIV Vif hijacking Cullin RING E3 ligase 5 is essential for HIV proliferation. H{\"u}ttenhain et al. discovered the host factor ARIH2, which works with CUL5Vif/CBF{\ss} via a tag-team mechanism to target APOBEC3 proteins for ubiquitination and is essential for HIV proliferation in primary CD4+ T cells.",
keywords = "AP-MS, APOBEC3, ARIH2, CUL5, HIV, Vif, host-virus interactions",
author = "Ruth H{\"u}ttenhain and Jiewei Xu and Burton, {Lily A.} and Gordon, {David E.} and Hultquist, {Judd F.} and Johnson, {Jeffrey R.} and L. Satkamp and Joseph Hiatt and Rhee, {David Y.} and Kheewoong Baek and Crosby, {David C.} and Frankel, {Alan D.} and Alexander Marson and Harper, {J. Wade} and Alpi, {Arno F.} and Schulman, {Brenda A.} and Gross, {John D.} and Krogan, {Nevan J.}",
note = "Funding Information: R.H. is a recipient of postdoctoral fellowships from the Swiss National Science Foundation ( P2EZP3_148742 ; P300P3_151154 ), the European Molecular Biology Organization ( ALTF 1123-2013 ), and the Human Frontiers in Science Program ( LT000089/2014-L ). R.H. was also supported by NIH funding for the UCSF-Gladstone Institute of Virology and Immunology Center for AIDS Research (CFAR; P30-AI027763 ). J.D.G., A.D.F., L.A.B., and D.C.C. were supported by NIH grant P50 GM082250 . J.F.H. is supported by AmfAR grant 109504-61-RKRL with funds raised by generationCURE, NIH grant K22 AI136691 , and a supplement from the NIH-supported Third Coast CFAR SP0029591 . N.J.K. was supported by NIH grants P50 GM082250 , U19 AI106754 , P01 HL089707 , P01 CA177332 , U19 AI118610 , R01 AI120694 , and P01 AI063302 . J.W.H. was supported by RO1 AG011085 . B.A.S. was supported by R37GM069530 , ALSAC /St. Jude, and the Max Planck Society . A.M. was supported in part by the NIH ( NIGMS P50 GM082250 and NIDA DP2 DA042423 ). A.M. holds a Career Award for Medical Scientists from the Burroughs Wellcome Fund, is an investigator at the Chan Zuckerberg Biohub, has received funding from the Innovative Genomics Institute (IGI), and is a member of the Parker Institute for Cancer Immunotherapy (PICI). The Marson lab has received a gift from Gilead to support HIV cure research. The proteomics work was carried out in the Thermo Fisher Scientific Mass Spectrometry Facility for Disease Target Discovery at the J. David Gladstone Institutes. The virus experiments were performed in the BSL3 facility at the J. David Gladstone Institutes. We thank members of the Krogan lab for helpful advice and comments and Mike Shales for help with figure design. Funding Information: R.H. is a recipient of postdoctoral fellowships from the Swiss National Science Foundation (P2EZP3_148742; P300P3_151154), the European Molecular Biology Organization (ALTF 1123-2013), and the Human Frontiers in Science Program (LT000089/2014-L). R.H. was also supported by NIH funding for the UCSF-Gladstone Institute of Virology and Immunology Center for AIDS Research (CFAR; P30-AI027763). J.D.G. A.D.F. L.A.B. and D.C.C. were supported by NIH grant P50 GM082250. J.F.H. is supported by AmfAR grant 109504-61-RKRL with funds raised by generationCURE, NIH grant K22 AI136691, and a supplement from the NIH-supported Third Coast CFAR SP0029591. N.J.K. was supported by NIH grants P50 GM082250, U19 AI106754, P01 HL089707, P01 CA177332, U19 AI118610, R01 AI120694, and P01 AI063302. J.W.H. was supported by RO1 AG011085. B.A.S. was supported by R37GM069530, ALSAC/St. Jude, and the Max Planck Society. A.M. was supported in part by the NIH (NIGMS P50 GM082250 and NIDA DP2 DA042423). A.M. holds a Career Award for Medical Scientists from the Burroughs Wellcome Fund, is an investigator at the Chan Zuckerberg Biohub, has received funding from the Innovative Genomics Institute (IGI), and is a member of the Parker Institute for Cancer Immunotherapy (PICI). The Marson lab has received a gift from Gilead to support HIV cure research. The proteomics work was carried out in the Thermo Fisher Scientific Mass Spectrometry Facility for Disease Target Discovery at the J. David Gladstone Institutes. The virus experiments were performed in the BSL3 facility at the J. David Gladstone Institutes. We thank members of the Krogan lab for helpful advice and comments and Mike Shales for help with figure design. R.H. and N.J.K. designed the study. R.H. J.X. L.S. and D.C.C. cloned constructs for affinity purifications and CRISPR knockouts and generated and characterized stable cell lines. R.H. L.S. D.E.G. J.F.H. and J.H. generated and characterized virus for infections. R.H. performed affinity purifications of CUL5, CBF{\ss}, and ELOB in the context of infection. R.H. and J.R.J. performed global mass spectrometric analysis of CUL5, CBF{\ss}, and ELOB affinity purifications. R.H. analyzed proteomic data and performed protein-protein interaction scoring. R.H. performed targeted proteomic experiments and analyzed the resulting data. R.H. J.X. D.E.G. J.F.H. and J.H. performed experiments in primary T cells (CRISPR knockouts, HIV infections, FACS analysis, and data analysis). L.B. purified all proteins and performed all the in vitro ubiquitination assays for APOBEC3G. R.H. and J.X. performed the APOBEC3G packing assays in ARIH2 KO cell lines. D.Y.R. performed affinity purifications, MS and data analysis for ARIH2. K.B. performed in vitro ubiquitination assays for CKB. R.H. A.M. A.D.F. J.W.H. A.F.A. B.A.S. J.D.G. and N.J.K. supervised research. R.H. and N.J.K. wrote the manuscript with input from B.A.S. and J.D.G. A.M. is a co-founder of Arsenal Biosciences, Spotlight Therapeutics, and Sonoma Biotherapeutics. A.M. serves on the scientific advisory board of PACT Pharma and was a former advisor to Juno Therapeutics. The Marson laboratory has received sponsored research support from Juno Therapeutics, Epinomics, and Sanofi and a gift from Gilead. Funding Information: A.M. is a co-founder of Arsenal Biosciences, Spotlight Therapeutics, and Sonoma Biotherapeutics. A.M. serves on the scientific advisory board of PACT Pharma and was a former advisor to Juno Therapeutics. The Marson laboratory has received sponsored research support from Juno Therapeutics, Epinomics, and Sanofi and a gift from Gilead. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
month = jul,
day = "10",
doi = "10.1016/j.chom.2019.05.008",
language = "English (US)",
volume = "26",
pages = "86--99.e7",
journal = "Cell Host and Microbe",
issn = "1931-3128",
publisher = "Cell Press",
number = "1",
}
