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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

SILAC-Based Proteomic Validation and Functional Analysis of Cell Surface Targets Downregulated by HIV-1(A) Workflow of SILAC-based 3-way PMP time course experiment.(B) Validation of HIV-1 targets (upper panel) and comparison between SILAC- and TMT-based time course experiments (lower panel). Log2(fold change compared with mock/uninfected cells) at 24, 48, and 72 hr is shown for proteins from Cluster #35 (red) versus all other quantitated proteins (gray). Downregulation by HIV-1 is indicated by dotted arrows. Proteins identified by >1 unique peptide in both TMT and SILAC experiments are shown. Crosses indicate mean values. ∗p < 0.05; ∗∗∗p < 0.001.(C) Gene ontology “molecular function” and “biological process” terms enriched among proteins from Cluster #35. DAVID functional annotation clusters with adjusted p values < 0.05 and containing terms with Bonferroni-adjusted p values < 0.05 are shown. Further details are included in Table S3.(D–F) Temporal profiles of downregulated proteins associated with cell adhesion (D), leukocyte activation (E), and transmembrane transport (F). Proteomic quantitation and time points are as for Figures 1B–1C. Proteins exhibiting >2-fold downregulation compared with uninfected cells in both TMT and SILAC experiments are shown, and proteins subsequently validated using flow cytometry or immunoblot are underlined.See also Figure S2 and Table S2.
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fig2: SILAC-Based Proteomic Validation and Functional Analysis of Cell Surface Targets Downregulated by HIV-1(A) Workflow of SILAC-based 3-way PMP time course experiment.(B) Validation of HIV-1 targets (upper panel) and comparison between SILAC- and TMT-based time course experiments (lower panel). Log2(fold change compared with mock/uninfected cells) at 24, 48, and 72 hr is shown for proteins from Cluster #35 (red) versus all other quantitated proteins (gray). Downregulation by HIV-1 is indicated by dotted arrows. Proteins identified by >1 unique peptide in both TMT and SILAC experiments are shown. Crosses indicate mean values. ∗p < 0.05; ∗∗∗p < 0.001.(C) Gene ontology “molecular function” and “biological process” terms enriched among proteins from Cluster #35. DAVID functional annotation clusters with adjusted p values < 0.05 and containing terms with Bonferroni-adjusted p values < 0.05 are shown. Further details are included in Table S3.(D–F) Temporal profiles of downregulated proteins associated with cell adhesion (D), leukocyte activation (E), and transmembrane transport (F). Proteomic quantitation and time points are as for Figures 1B–1C. Proteins exhibiting >2-fold downregulation compared with uninfected cells in both TMT and SILAC experiments are shown, and proteins subsequently validated using flow cytometry or immunoblot are underlined.See also Figure S2 and Table S2.

Mentions: We validated these candidates in an independent infection time course experiment using SILAC as an alternative quantitative proteomic approach (Figures 2A and 2B) and confirmed downregulation of a functionally and structurally diverse set of proteins with available antibody reagents (CD43/47/162 and NOTCH1) by flow cytometry in CEM-T4s and primary human CD4+ T cells infected with HIV-1 (Figure S2A).


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)

SILAC-Based Proteomic Validation and Functional Analysis of Cell Surface Targets Downregulated by HIV-1(A) Workflow of SILAC-based 3-way PMP time course experiment.(B) Validation of HIV-1 targets (upper panel) and comparison between SILAC- and TMT-based time course experiments (lower panel). Log2(fold change compared with mock/uninfected cells) at 24, 48, and 72 hr is shown for proteins from Cluster #35 (red) versus all other quantitated proteins (gray). Downregulation by HIV-1 is indicated by dotted arrows. Proteins identified by >1 unique peptide in both TMT and SILAC experiments are shown. Crosses indicate mean values. ∗p < 0.05; ∗∗∗p < 0.001.(C) Gene ontology “molecular function” and “biological process” terms enriched among proteins from Cluster #35. DAVID functional annotation clusters with adjusted p values < 0.05 and containing terms with Bonferroni-adjusted p values < 0.05 are shown. Further details are included in Table S3.(D–F) Temporal profiles of downregulated proteins associated with cell adhesion (D), leukocyte activation (E), and transmembrane transport (F). Proteomic quantitation and time points are as for Figures 1B–1C. Proteins exhibiting >2-fold downregulation compared with uninfected cells in both TMT and SILAC experiments are shown, and proteins subsequently validated using flow cytometry or immunoblot are underlined.See also Figure S2 and Table S2.
© Copyright Policy - CC BY
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4608997&req=5

fig2: SILAC-Based Proteomic Validation and Functional Analysis of Cell Surface Targets Downregulated by HIV-1(A) Workflow of SILAC-based 3-way PMP time course experiment.(B) Validation of HIV-1 targets (upper panel) and comparison between SILAC- and TMT-based time course experiments (lower panel). Log2(fold change compared with mock/uninfected cells) at 24, 48, and 72 hr is shown for proteins from Cluster #35 (red) versus all other quantitated proteins (gray). Downregulation by HIV-1 is indicated by dotted arrows. Proteins identified by >1 unique peptide in both TMT and SILAC experiments are shown. Crosses indicate mean values. ∗p < 0.05; ∗∗∗p < 0.001.(C) Gene ontology “molecular function” and “biological process” terms enriched among proteins from Cluster #35. DAVID functional annotation clusters with adjusted p values < 0.05 and containing terms with Bonferroni-adjusted p values < 0.05 are shown. Further details are included in Table S3.(D–F) Temporal profiles of downregulated proteins associated with cell adhesion (D), leukocyte activation (E), and transmembrane transport (F). Proteomic quantitation and time points are as for Figures 1B–1C. Proteins exhibiting >2-fold downregulation compared with uninfected cells in both TMT and SILAC experiments are shown, and proteins subsequently validated using flow cytometry or immunoblot are underlined.See also Figure S2 and Table S2.
Mentions: We validated these candidates in an independent infection time course experiment using SILAC as an alternative quantitative proteomic approach (Figures 2A and 2B) and confirmed downregulation of a functionally and structurally diverse set of proteins with available antibody reagents (CD43/47/162 and NOTCH1) by flow cytometry in CEM-T4s and primary human CD4+ T cells infected with HIV-1 (Figure S2A).

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