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Cell Surface Proteomic Map of HIV Infection Reveals Antagonism of Amino Acid Metabolism by Vpu and Nef.

Matheson NJ, Sumner J, Wals K, Rapiteanu R, Weekes MP, Vigan R, Weinelt J, Schindler M, Antrobus R, Costa AS, Frezza C, Clish CB, Neil SJ, Lehner PJ - Cell Host Microbe (2015)

Bottom Line: SNAT1 antagonism was acquired by Vpu variants from the lineage of SIVcpz/HIV-1 viruses responsible for pandemic AIDS.We found marked SNAT1 induction in activated primary human CD4+ T cells, and used Consumption and Release (CoRe) metabolomics to identify alanine as an endogenous SNAT1 substrate required for T cell mitogenesis.Downregulation of SNAT1 therefore defines a unique paradigm of HIV interference with immunometabolism.

View Article: PubMed Central - PubMed

Affiliation: Cambridge Institute for Medical Research, University of Cambridge, Cambridge Biomedical Campus, Cambridge CB2 0XY, UK. Electronic address: njm25@cam.ac.uk.

No MeSH data available.


Related in: MedlinePlus

SNAT1 Downregulation by Vpu Variants from Pandemic HIV-1 Viruses(A) Screening strategy for SNAT1 downregulation by naturally occurring Vpu variants. 293Ts stably expressing SNAT1-FLAG and CD4 were transfected with the indicated pCG-IRES-GFP constructs (all based on HIV-1 group M, clade B, strain NL4-3 virus) and analyzed by flow cytometry at 36 hr. Target downregulation is indicated by a shift in the transfected (GFP+) cells toward the lower left quadrant (red arrows).(B) SNAT1-FLAG downregulation by Vpu variants from pandemic HIV-1 group M clade A/B/C viruses. As for (A), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1.(C) Phylogenetic analysis of SNAT1-FLAG downregulation by Vpu variants of HIV-1 and SIV viruses. As for (A) and (B), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1 or SIV and downregulation of SNAT1-FLAG or CD4 expressed as ratio of geomean fluorescence intensity between transfected (GFP+) and untransfected (GFP−) cells. Illustrative phylogenetic relationships are shown, and branch lengths are arbitrary (further details are included in Supplemental Experimental Procedures). HIV-1/M/N/O (HIV-1 group M, N, or O viruses); SIVcpz Ptt (SIVs infecting central P. t. troglodytes chimpanzees); SIVcpz Pts (SIVs infecting eastern P. t. schweinfruthii chimpanzees); SIVgor (gorilla SIV); SIVguenon (SIVs infecting guenon monkeys).
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fig7: SNAT1 Downregulation by Vpu Variants from Pandemic HIV-1 Viruses(A) Screening strategy for SNAT1 downregulation by naturally occurring Vpu variants. 293Ts stably expressing SNAT1-FLAG and CD4 were transfected with the indicated pCG-IRES-GFP constructs (all based on HIV-1 group M, clade B, strain NL4-3 virus) and analyzed by flow cytometry at 36 hr. Target downregulation is indicated by a shift in the transfected (GFP+) cells toward the lower left quadrant (red arrows).(B) SNAT1-FLAG downregulation by Vpu variants from pandemic HIV-1 group M clade A/B/C viruses. As for (A), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1.(C) Phylogenetic analysis of SNAT1-FLAG downregulation by Vpu variants of HIV-1 and SIV viruses. As for (A) and (B), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1 or SIV and downregulation of SNAT1-FLAG or CD4 expressed as ratio of geomean fluorescence intensity between transfected (GFP+) and untransfected (GFP−) cells. Illustrative phylogenetic relationships are shown, and branch lengths are arbitrary (further details are included in Supplemental Experimental Procedures). HIV-1/M/N/O (HIV-1 group M, N, or O viruses); SIVcpz Ptt (SIVs infecting central P. t. troglodytes chimpanzees); SIVcpz Pts (SIVs infecting eastern P. t. schweinfruthii chimpanzees); SIVgor (gorilla SIV); SIVguenon (SIVs infecting guenon monkeys).

Mentions: To explore the phylogenetic history of Vpu-mediated SNAT1 downregulation, we generated a stable 293T cell line expressing SNAT1-FLAG and CD4 and compared the effects of different Vpu-IRES-GFP constructs. CD4 downregulation is widely conserved and therefore represents a positive control for functional Vpu expression (Sauter et al., 2009). As expected, whereas NL4-3 Vpu (but not Vpu S52A) downregulated both CD4 and SNAT1-FLAG, Nef only downregulated CD4 (Figure 7A). Depletion of cell surface SNAT1-FLAG was conserved across all 6 HIV-1 clade A, B, and C Vpu variants tested (Figures 7A and 7B), but restricted to the SIVcpz Ptt lineage giving rise to pandemic HIV-1 group M viruses (Figure 7C). The ability of Vpu to downregulate SNAT1 has therefore been acquired recently and may be critical for the in vivo replication or enhanced pathogenicity of HIV-1 viruses.


Cell Surface Proteomic Map of HIV Infection Reveals Antagonism of Amino Acid Metabolism by Vpu and Nef.

Matheson NJ, Sumner J, Wals K, Rapiteanu R, Weekes MP, Vigan R, Weinelt J, Schindler M, Antrobus R, Costa AS, Frezza C, Clish CB, Neil SJ, Lehner PJ - Cell Host Microbe (2015)

SNAT1 Downregulation by Vpu Variants from Pandemic HIV-1 Viruses(A) Screening strategy for SNAT1 downregulation by naturally occurring Vpu variants. 293Ts stably expressing SNAT1-FLAG and CD4 were transfected with the indicated pCG-IRES-GFP constructs (all based on HIV-1 group M, clade B, strain NL4-3 virus) and analyzed by flow cytometry at 36 hr. Target downregulation is indicated by a shift in the transfected (GFP+) cells toward the lower left quadrant (red arrows).(B) SNAT1-FLAG downregulation by Vpu variants from pandemic HIV-1 group M clade A/B/C viruses. As for (A), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1.(C) Phylogenetic analysis of SNAT1-FLAG downregulation by Vpu variants of HIV-1 and SIV viruses. As for (A) and (B), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1 or SIV and downregulation of SNAT1-FLAG or CD4 expressed as ratio of geomean fluorescence intensity between transfected (GFP+) and untransfected (GFP−) cells. Illustrative phylogenetic relationships are shown, and branch lengths are arbitrary (further details are included in Supplemental Experimental Procedures). HIV-1/M/N/O (HIV-1 group M, N, or O viruses); SIVcpz Ptt (SIVs infecting central P. t. troglodytes chimpanzees); SIVcpz Pts (SIVs infecting eastern P. t. schweinfruthii chimpanzees); SIVgor (gorilla SIV); SIVguenon (SIVs infecting guenon monkeys).
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fig7: SNAT1 Downregulation by Vpu Variants from Pandemic HIV-1 Viruses(A) Screening strategy for SNAT1 downregulation by naturally occurring Vpu variants. 293Ts stably expressing SNAT1-FLAG and CD4 were transfected with the indicated pCG-IRES-GFP constructs (all based on HIV-1 group M, clade B, strain NL4-3 virus) and analyzed by flow cytometry at 36 hr. Target downregulation is indicated by a shift in the transfected (GFP+) cells toward the lower left quadrant (red arrows).(B) SNAT1-FLAG downregulation by Vpu variants from pandemic HIV-1 group M clade A/B/C viruses. As for (A), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1.(C) Phylogenetic analysis of SNAT1-FLAG downregulation by Vpu variants of HIV-1 and SIV viruses. As for (A) and (B), but cells were transfected with pCG-IRES-GFP constructs encoding Vpu variants from the indicated strains of HIV-1 or SIV and downregulation of SNAT1-FLAG or CD4 expressed as ratio of geomean fluorescence intensity between transfected (GFP+) and untransfected (GFP−) cells. Illustrative phylogenetic relationships are shown, and branch lengths are arbitrary (further details are included in Supplemental Experimental Procedures). HIV-1/M/N/O (HIV-1 group M, N, or O viruses); SIVcpz Ptt (SIVs infecting central P. t. troglodytes chimpanzees); SIVcpz Pts (SIVs infecting eastern P. t. schweinfruthii chimpanzees); SIVgor (gorilla SIV); SIVguenon (SIVs infecting guenon monkeys).
Mentions: To explore the phylogenetic history of Vpu-mediated SNAT1 downregulation, we generated a stable 293T cell line expressing SNAT1-FLAG and CD4 and compared the effects of different Vpu-IRES-GFP constructs. CD4 downregulation is widely conserved and therefore represents a positive control for functional Vpu expression (Sauter et al., 2009). As expected, whereas NL4-3 Vpu (but not Vpu S52A) downregulated both CD4 and SNAT1-FLAG, Nef only downregulated CD4 (Figure 7A). Depletion of cell surface SNAT1-FLAG was conserved across all 6 HIV-1 clade A, B, and C Vpu variants tested (Figures 7A and 7B), but restricted to the SIVcpz Ptt lineage giving rise to pandemic HIV-1 group M viruses (Figure 7C). The ability of Vpu to downregulate SNAT1 has therefore been acquired recently and may be critical for the in vivo replication or enhanced pathogenicity of HIV-1 viruses.

Bottom Line: SNAT1 antagonism was acquired by Vpu variants from the lineage of SIVcpz/HIV-1 viruses responsible for pandemic AIDS.We found marked SNAT1 induction in activated primary human CD4+ T cells, and used Consumption and Release (CoRe) metabolomics to identify alanine as an endogenous SNAT1 substrate required for T cell mitogenesis.Downregulation of SNAT1 therefore defines a unique paradigm of HIV interference with immunometabolism.

View Article: PubMed Central - PubMed

Affiliation: Cambridge Institute for Medical Research, University of Cambridge, Cambridge Biomedical Campus, Cambridge CB2 0XY, UK. Electronic address: njm25@cam.ac.uk.

No MeSH data available.


Related in: MedlinePlus