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Pulsed stable isotope labeling of amino acids in cell culture uncovers the dynamic interactions between HIV-1 and the monocyte-derived macrophage.

Kraft-Terry SD, Engebretsen IL, Bastola DK, Fox HS, Ciborowski P, Gendelman HE - J. Proteome Res. (2011)

Bottom Line: Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased.Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication.We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska 68198-5880, USA.

ABSTRACT
Dynamic interactions between human immunodeficiency virus-1 (HIV-1) and the macrophage govern the tempo of viral dissemination and replication in its human host. HIV-1 affects macrophage phenotype, and the macrophage, in turn, can modulate the viral life cycle. While these processes are linked to host-cell function and survival, the precise intracellular pathways involved are incompletely understood. To elucidate such dynamic virus-cell events, we employed pulsed stable isotope labeling of amino acids in cell culture. Alterations in de novo protein synthesis of HIV-1 infected human monocyte-derived macrophages (MDM) were examined after 3, 5, and 7 days of viral infection. Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased. Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication. We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

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(A) Venn diagram of the overlap in protein identification with significant ratios (Significance B ≤ 0.05) between days 3, 5, and 7. Distribution of log2 transformed protein expression ratios of HIV-1 to control on (B) day 3, (C) day 5, and (D) day 7 post HIV-1 infection. Ratios are graphed versus intensity.
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fig3: (A) Venn diagram of the overlap in protein identification with significant ratios (Significance B ≤ 0.05) between days 3, 5, and 7. Distribution of log2 transformed protein expression ratios of HIV-1 to control on (B) day 3, (C) day 5, and (D) day 7 post HIV-1 infection. Ratios are graphed versus intensity.

Mentions: A total of 393 proteins were identified as differentially expressed, with significance values of ≤0.05 for at least one of the 3 time points examined after HIV-1 infection (Figure 3a). At all time points, 28 proteins were identified as differentially expressed. The number of proteins that were identified between only two of the time points were similar in quantity, no matter which overlapping days were examined. Thirty-two, 28, and 24 common proteins with significant ratios between infected and uninfected MDM were observed on days 3 and 5, 5 and 7, or 3 and 7 after infection, respectively. The numbers of unique proteins with differentially expressed synthesis rates at single time points were 99, 77, and 105 on days 3, 5, and 7 after infection, respectively. These differences demonstrate that the cellular response to HIV-1 infection change during the course of viral infection.


Pulsed stable isotope labeling of amino acids in cell culture uncovers the dynamic interactions between HIV-1 and the monocyte-derived macrophage.

Kraft-Terry SD, Engebretsen IL, Bastola DK, Fox HS, Ciborowski P, Gendelman HE - J. Proteome Res. (2011)

(A) Venn diagram of the overlap in protein identification with significant ratios (Significance B ≤ 0.05) between days 3, 5, and 7. Distribution of log2 transformed protein expression ratios of HIV-1 to control on (B) day 3, (C) day 5, and (D) day 7 post HIV-1 infection. Ratios are graphed versus intensity.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108467&req=5

fig3: (A) Venn diagram of the overlap in protein identification with significant ratios (Significance B ≤ 0.05) between days 3, 5, and 7. Distribution of log2 transformed protein expression ratios of HIV-1 to control on (B) day 3, (C) day 5, and (D) day 7 post HIV-1 infection. Ratios are graphed versus intensity.
Mentions: A total of 393 proteins were identified as differentially expressed, with significance values of ≤0.05 for at least one of the 3 time points examined after HIV-1 infection (Figure 3a). At all time points, 28 proteins were identified as differentially expressed. The number of proteins that were identified between only two of the time points were similar in quantity, no matter which overlapping days were examined. Thirty-two, 28, and 24 common proteins with significant ratios between infected and uninfected MDM were observed on days 3 and 5, 5 and 7, or 3 and 7 after infection, respectively. The numbers of unique proteins with differentially expressed synthesis rates at single time points were 99, 77, and 105 on days 3, 5, and 7 after infection, respectively. These differences demonstrate that the cellular response to HIV-1 infection change during the course of viral infection.

Bottom Line: Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased.Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication.We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska 68198-5880, USA.

ABSTRACT
Dynamic interactions between human immunodeficiency virus-1 (HIV-1) and the macrophage govern the tempo of viral dissemination and replication in its human host. HIV-1 affects macrophage phenotype, and the macrophage, in turn, can modulate the viral life cycle. While these processes are linked to host-cell function and survival, the precise intracellular pathways involved are incompletely understood. To elucidate such dynamic virus-cell events, we employed pulsed stable isotope labeling of amino acids in cell culture. Alterations in de novo protein synthesis of HIV-1 infected human monocyte-derived macrophages (MDM) were examined after 3, 5, and 7 days of viral infection. Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased. Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication. We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

Show MeSH
Related in: MedlinePlus