Abstract
In contrast to activated CD4+ T cells, resting human CD4+ T cells circulating in blood are highly resistant to infection with human immunodeficiency virus (HIV)1,2,3,4. Whether the inability of HIV to infect these resting CD4+ T cells is due to the lack of a key factor, or alternatively reflects the presence of an efficient mechanism for defence against HIV, is not clear. Here we show that the anti-retroviral deoxycytidine deaminase APOBEC3G5 strongly protects unstimulated peripheral blood CD4+ T cells against HIV-1 infection. In activated CD4+ T cells, cytoplasmic APOBEC3G resides in an enzymatically inactive, high-molecular-mass (HMM) ribonucleoprotein complex that converts to an enzymatically active low-molecular-mass (LMM) form after treatment with RNase. In contrast, LMM APOBEC3G predominates in unstimulated CD4+ T cells, where HIV-1 replication is blocked and reverse transcription is impaired1,2,3. Mitogen activation induces the recruitment of LMM APOBEC3G into the HMM complex, and this correlates with a sharp increase in permissivity for HIV infection in these stimulated cells. Notably, when APOBEC3G-specific small interfering RNAs are introduced into unstimulated CD4+ T cells, the early replication block encountered by HIV-1 is greatly relieved. Thus, LMM APOBEC3G functions as a potent post-entry restriction factor for HIV-1 in unstimulated CD4+ T cells. Surprisingly, sequencing of the reverse transcripts slowly formed in unstimulated CD4+ T cells reveals only low levels of dG → dA hypermutation, raising the possibility that the APOBEC3G-restricting activity may not be strictly dependent on deoxycytidine deamination.
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Acknowledgements
We thank K. Strebel for the gift of the pNL-A1 plasmid, X.-F. Yu for the gift of the Myc-Cul5 plasmid, D. Bohmann for the gift of the His6-tagged ubiquitin expression vector DNA, N. R. Landau for the gift of NL4-3 HSA R-E- reporter provirus through the AIDS Research and Reference Reagent Program at NIH, J. Burns for the gift of expression plasmid pVSV-G, and M. Malim for providing the monoclonal anti-Vif (319) antibody. We thank A. O'Mahony and H. Kwon for assistance with the FPLC experiments; J. Neidleman for assistance preparing the primary cells; M. Cavrois, D. Fenard, A. Yonezawa, J. Bohuslav, L.-F. Chen, C. Martin and S. Williams for discussions; G. Howard and S. Ordway for editorial assistance; and S. Cammack, R. Givens and J. Carroll for assistance in preparation of the manuscript and the graphics. Different components of this work were supported by funding from the National Institutes of Health (Women's HIV Interdisciplinary Network and NIMH; W.C.G.) and the Universitywide AIDS Research Program (W.C.G.) and the American Foundation for AIDS Research (Y.-L.C.).
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Supplementary information
Supplementary Figure S1
Enzymatic activity of RNase removes RNA molecules associated with the HMW A3G complexes and promotes the disassembly of these complexes. (PDF 46 kb)
Supplementary Figure S2
Vif recruits Cul5 and elongin B, components of an ECS (elongin B-Cul5-SOCS) ubiquitin ligase complex to the HMW HA-A3G complex. (PDF 67 kb)
Supplementary Figure 3
Permissiveness of CD4 T-cells to HIV infection before or after stimulation. (PDF 125 kb)
Supplementary Table S1
The primer and probe sets for real-time PCR and sequencing analysis of HIV-1 reverse transcripts. (DOC 21 kb)
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Chiu, YL., Soros, V., Kreisberg, J. et al. Cellular APOBEC3G restricts HIV-1 infection in resting CD4+ T cells. Nature 435, 108–114 (2005). https://doi.org/10.1038/nature03493
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DOI: https://doi.org/10.1038/nature03493
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