The Role of RNA in HIV-1 Vif-Mediated Degradation of APOBEC3H

Jiayi Wang, Jordan T. Becker, Ke Shi, Kate V. Lauer, Daniel J. Salamango, Hideki Aihara, Nadine M. Shaban, Reuben S. Harris

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

As many as five members of the APOBEC3 family of DNA cytosine deaminases are capable of inhibiting HIV-1 replication by deaminating viral cDNA cytosines and interfering with reverse transcription. HIV-1 counteracts restriction with the virally encoded Vif protein, which forms a hybrid ubiquitin ligase complex that directly binds APOBEC3 enzymes and targets them for proteasomal degradation. APOBEC3H (A3H) is unique among family members by dimerization through cellular and viral duplex RNA species. RNA binding is required for localization of A3H to the cytoplasmic compartment, for efficient packaging into nascent HIV-1 particles and ultimately for effective virus restriction activity. Here we compared wild-type human A3H and RNA binding–defective mutants to ask whether RNA may be a factor in the functional interaction with HIV-1 Vif. We used structural modeling, immunoblotting, live cell imaging, and split green fluorescence protein (GFP) reconstitution approaches to assess the capability of HIV-1 Vif to promote the degradation of wild-type A3H in comparison to RNA binding–defective mutants. The results combined to show that RNA is not strictly required for Vif-mediated degradation of A3H, and that RNA and Vif are likely to bind this single-domain DNA cytosine deaminase on physically distinct surfaces. However, a subset of the results also indicated that the A3H degradation process may be affected by A3H protein structure, subcellular localization, and differences in the constellation of A3H interaction partners, suggesting additional factors may also influence the fate and functionality of this host-pathogen interaction.

Original languageEnglish (US)
Pages (from-to)5019-5031
Number of pages13
JournalJournal of Molecular Biology
Volume431
Issue number24
DOIs
StatePublished - Dec 6 2019

Bibliographical note

Funding Information:
This work was supported by NIAID R37 AI064046 (to R.S.H.), NIGMS R01 GM118000 (to R.S.H. and H.A.), and NIGMS R01 GM118047 (to H.A.). J.W. received partial salary support from the University of Minnesota Graduate School , Interdisciplinary Doctoral Fellowship. D.J.S. received partial salary support from the University of Minnesota Craniofacial Research Training (MinnCResT) program ( NIH T90DE022732 ). R.S.H. is the Margaret Harvey Schering Land Grant Chair for Cancer Research, a Distinguished University McKnight Professor, and an Investigator of the Howard Hughes Medical Institute. The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pRGH-WT (#12427) from Ivan Sadowski and Viviana Simon. Microscopy imaging and analysis was performed at the University Imaging Centers, University of Minnesota.

Funding Information:
This work was supported by NIAID R37 AI064046 (to R.S.H.), NIGMS R01 GM118000 (to R.S.H. and H.A.), and NIGMS R01 GM118047 (to H.A.). J.W. received partial salary support from the University of Minnesota Graduate School, Interdisciplinary Doctoral Fellowship. D.J.S. received partial salary support from the University of Minnesota Craniofacial Research Training (MinnCResT) program (NIH T90DE022732). R.S.H. is the Margaret Harvey Schering Land Grant Chair for Cancer Research, a Distinguished University McKnight Professor, and an Investigator of the Howard Hughes Medical Institute. The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pRGH-WT (#12427) from Ivan Sadowski and Viviana Simon. Microscopy imaging and analysis was performed at the University Imaging Centers, University of Minnesota.

Keywords

  • APOBEC3H
  • DNA cytosine deaminase
  • HIV-1 Vif
  • Subcellular localization
  • Vif-APOBEC interaction

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