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Global Reprogramming of Host SUMOylation during Influenza Virus Infection.

Domingues P, Golebiowski F, Tatham MH, Lopes AM, Taggart A, Hay RT, Hale BG - Cell Rep (2015)

Bottom Line: This is paralleled by widespread host deSUMOylation.Depletion screening identified ten virus-induced SUMO targets as potential antiviral factors, including C18orf25 and the SMC5/6 and PAF1 complexes.Mechanistic studies further uncovered a role for SUMOylation of the PAF1 complex component, parafibromin (CDC73), in potentiating antiviral gene expression.

View Article: PubMed Central - PubMed

Affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Campus, 464 Bearsden Road, Glasgow G61 1QH, UK.

No MeSH data available.


Related in: MedlinePlus

SUMO Targets Impacting IAV Replication(A) Schematic representation of the lentivirus-based shRNA screen assessing 42 host SUMO targets for their impact on IAV replication.(B) Heatmap summary of factors identified as required or restrictive to IAV replication in A549 cells. Genes are shown whose depletion led to a 5-fold or more difference in infectious IAV titer as compared with control for at least two of three shRNA sequences. Each individual shRNA is labeled a, b, or c and control shRNAs are highlighted in gray.(C–E) Validation of PAF1 (C), C18orf25 (D), and AFF4 (E) as impacting IAV replication. The two shRNA sequences for each gene from (B) that showed consistent impact on IAV replication were independently validated in the same assay for their effect on IAV replication (top) and specific gene knockdown and effect on cell viability (bottom). Bars represent mean values from triplicates (±SD). See also Table S6.
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fig6: SUMO Targets Impacting IAV Replication(A) Schematic representation of the lentivirus-based shRNA screen assessing 42 host SUMO targets for their impact on IAV replication.(B) Heatmap summary of factors identified as required or restrictive to IAV replication in A549 cells. Genes are shown whose depletion led to a 5-fold or more difference in infectious IAV titer as compared with control for at least two of three shRNA sequences. Each individual shRNA is labeled a, b, or c and control shRNAs are highlighted in gray.(C–E) Validation of PAF1 (C), C18orf25 (D), and AFF4 (E) as impacting IAV replication. The two shRNA sequences for each gene from (B) that showed consistent impact on IAV replication were independently validated in the same assay for their effect on IAV replication (top) and specific gene knockdown and effect on cell viability (bottom). Bars represent mean values from triplicates (±SD). See also Table S6.

Mentions: Consistent between our SUMO1 and SUMO2 proteomic studies, IAV infection triggered a >4-fold increase in SUMOylation of 42 host proteins. To identify functional roles for these core SUMO targets during IAV infection, we depleted A549 cells of the corresponding 42 genes one by one using small hairpin RNA (shRNA)-expressing lentiviruses (three per gene), and we determined the subsequent replication of IAV by measuring infectious virus yields at 24 and 48 hr post-infection (Figure 6A). As controls, we also assessed the impact on IAV replication of depleting IRF3, a host antiviral defense transcription factor, and ATP6V0C, a vacuolar ATPase component required for efficient IAV entry (König et al., 2010). We classified a host gene as important for IAV replication if at least two of three shRNAs increased or decreased infectious IAV yields by at least 5-fold at a minimum of one time point compared to the non-targeting shRNA.


Global Reprogramming of Host SUMOylation during Influenza Virus Infection.

Domingues P, Golebiowski F, Tatham MH, Lopes AM, Taggart A, Hay RT, Hale BG - Cell Rep (2015)

SUMO Targets Impacting IAV Replication(A) Schematic representation of the lentivirus-based shRNA screen assessing 42 host SUMO targets for their impact on IAV replication.(B) Heatmap summary of factors identified as required or restrictive to IAV replication in A549 cells. Genes are shown whose depletion led to a 5-fold or more difference in infectious IAV titer as compared with control for at least two of three shRNA sequences. Each individual shRNA is labeled a, b, or c and control shRNAs are highlighted in gray.(C–E) Validation of PAF1 (C), C18orf25 (D), and AFF4 (E) as impacting IAV replication. The two shRNA sequences for each gene from (B) that showed consistent impact on IAV replication were independently validated in the same assay for their effect on IAV replication (top) and specific gene knockdown and effect on cell viability (bottom). Bars represent mean values from triplicates (±SD). See also Table S6.
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fig6: SUMO Targets Impacting IAV Replication(A) Schematic representation of the lentivirus-based shRNA screen assessing 42 host SUMO targets for their impact on IAV replication.(B) Heatmap summary of factors identified as required or restrictive to IAV replication in A549 cells. Genes are shown whose depletion led to a 5-fold or more difference in infectious IAV titer as compared with control for at least two of three shRNA sequences. Each individual shRNA is labeled a, b, or c and control shRNAs are highlighted in gray.(C–E) Validation of PAF1 (C), C18orf25 (D), and AFF4 (E) as impacting IAV replication. The two shRNA sequences for each gene from (B) that showed consistent impact on IAV replication were independently validated in the same assay for their effect on IAV replication (top) and specific gene knockdown and effect on cell viability (bottom). Bars represent mean values from triplicates (±SD). See also Table S6.
Mentions: Consistent between our SUMO1 and SUMO2 proteomic studies, IAV infection triggered a >4-fold increase in SUMOylation of 42 host proteins. To identify functional roles for these core SUMO targets during IAV infection, we depleted A549 cells of the corresponding 42 genes one by one using small hairpin RNA (shRNA)-expressing lentiviruses (three per gene), and we determined the subsequent replication of IAV by measuring infectious virus yields at 24 and 48 hr post-infection (Figure 6A). As controls, we also assessed the impact on IAV replication of depleting IRF3, a host antiviral defense transcription factor, and ATP6V0C, a vacuolar ATPase component required for efficient IAV entry (König et al., 2010). We classified a host gene as important for IAV replication if at least two of three shRNAs increased or decreased infectious IAV yields by at least 5-fold at a minimum of one time point compared to the non-targeting shRNA.

Bottom Line: This is paralleled by widespread host deSUMOylation.Depletion screening identified ten virus-induced SUMO targets as potential antiviral factors, including C18orf25 and the SMC5/6 and PAF1 complexes.Mechanistic studies further uncovered a role for SUMOylation of the PAF1 complex component, parafibromin (CDC73), in potentiating antiviral gene expression.

View Article: PubMed Central - PubMed

Affiliation: MRC-University of Glasgow Centre for Virus Research, Garscube Campus, 464 Bearsden Road, Glasgow G61 1QH, UK.

No MeSH data available.


Related in: MedlinePlus