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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

Quantitative SUMO Proteomics of IAV-Infected Cells(A) SILAC-based SUMO1 and SUMO2 proteomic workflow. The specified A549s were grown for five to six cell doublings in light (L; isotopically normal, K0R0), medium (M; K4R6), or heavy (H; K8R10) SILAC medium prior to treatment and processing as indicated.(B) The tsMAPs of SUMO1 (left) and SUMO2 (right) substrates after data filtering, indicating log2-fold changes in protein modification following IAV infection (y axis). The numbers of substrates identified in each category are indicated and certain examples are highlighted.(C) Correlation of log2-fold changes in SUMO1 and SUMO2 substrate modification following IAV infection. The 63 substrates that increase (and 158 substrates that decrease) in both SUMO1 and SUMO2 modification following IAV infection are highlighted, and certain example proteins are labeled.(D) The 63 host substrates that increase in SUMOylation with IAV infection organized by manually curated functional category. See also Figure S3 and Tables S1, S2, S3, S4, and S7.
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fig3: Quantitative SUMO Proteomics of IAV-Infected Cells(A) SILAC-based SUMO1 and SUMO2 proteomic workflow. The specified A549s were grown for five to six cell doublings in light (L; isotopically normal, K0R0), medium (M; K4R6), or heavy (H; K8R10) SILAC medium prior to treatment and processing as indicated.(B) The tsMAPs of SUMO1 (left) and SUMO2 (right) substrates after data filtering, indicating log2-fold changes in protein modification following IAV infection (y axis). The numbers of substrates identified in each category are indicated and certain examples are highlighted.(C) Correlation of log2-fold changes in SUMO1 and SUMO2 substrate modification following IAV infection. The 63 substrates that increase (and 158 substrates that decrease) in both SUMO1 and SUMO2 modification following IAV infection are highlighted, and certain example proteins are labeled.(D) The 63 host substrates that increase in SUMOylation with IAV infection organized by manually curated functional category. See also Figure S3 and Tables S1, S2, S3, S4, and S7.

Mentions: We conducted two independent stable isotope labeling by amino acids in cell culture (SILAC) experiments to determine the impact of IAV infection for 10 hr on the SUMO1 and SUMO2 sub-proteomes of A549 cells (Figure 3A). This time point was chosen to ensure all infected cells had undergone a full single cycle of virus replication and to capture primary dynamic SUMOylation changes. Notably, <2% of cellular proteins varied in total abundance more than ∼2-fold either between the TAP-only and TAP-SUMO cell lines or after IAV infection (Figures S3D and S3E). In contrast, analysis of the purified samples showed that ∼32% (SUMO1) and ∼47% (SUMO2) of quantified proteins were >2-fold more abundant in the purified TAP-SUMO material compared with the purified TAP-only material, and ∼36% (SUMO1) and ∼25% (SUMO2) of quantified proteins varied >2-fold in abundance in the purified TAP-SUMO material following IAV infection (Figures S3F and S3G). Together, this suggests that a large proportion of identified and quantified proteins in the purified, but not crude, samples show specific changes in abundance relating to SUMO modification status, as well as a dependence upon infection for SUMO conjugation state.


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)

Quantitative SUMO Proteomics of IAV-Infected Cells(A) SILAC-based SUMO1 and SUMO2 proteomic workflow. The specified A549s were grown for five to six cell doublings in light (L; isotopically normal, K0R0), medium (M; K4R6), or heavy (H; K8R10) SILAC medium prior to treatment and processing as indicated.(B) The tsMAPs of SUMO1 (left) and SUMO2 (right) substrates after data filtering, indicating log2-fold changes in protein modification following IAV infection (y axis). The numbers of substrates identified in each category are indicated and certain examples are highlighted.(C) Correlation of log2-fold changes in SUMO1 and SUMO2 substrate modification following IAV infection. The 63 substrates that increase (and 158 substrates that decrease) in both SUMO1 and SUMO2 modification following IAV infection are highlighted, and certain example proteins are labeled.(D) The 63 host substrates that increase in SUMOylation with IAV infection organized by manually curated functional category. See also Figure S3 and Tables S1, S2, S3, S4, and S7.
© Copyright Policy - CC BY
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fig3: Quantitative SUMO Proteomics of IAV-Infected Cells(A) SILAC-based SUMO1 and SUMO2 proteomic workflow. The specified A549s were grown for five to six cell doublings in light (L; isotopically normal, K0R0), medium (M; K4R6), or heavy (H; K8R10) SILAC medium prior to treatment and processing as indicated.(B) The tsMAPs of SUMO1 (left) and SUMO2 (right) substrates after data filtering, indicating log2-fold changes in protein modification following IAV infection (y axis). The numbers of substrates identified in each category are indicated and certain examples are highlighted.(C) Correlation of log2-fold changes in SUMO1 and SUMO2 substrate modification following IAV infection. The 63 substrates that increase (and 158 substrates that decrease) in both SUMO1 and SUMO2 modification following IAV infection are highlighted, and certain example proteins are labeled.(D) The 63 host substrates that increase in SUMOylation with IAV infection organized by manually curated functional category. See also Figure S3 and Tables S1, S2, S3, S4, and S7.
Mentions: We conducted two independent stable isotope labeling by amino acids in cell culture (SILAC) experiments to determine the impact of IAV infection for 10 hr on the SUMO1 and SUMO2 sub-proteomes of A549 cells (Figure 3A). This time point was chosen to ensure all infected cells had undergone a full single cycle of virus replication and to capture primary dynamic SUMOylation changes. Notably, <2% of cellular proteins varied in total abundance more than ∼2-fold either between the TAP-only and TAP-SUMO cell lines or after IAV infection (Figures S3D and S3E). In contrast, analysis of the purified samples showed that ∼32% (SUMO1) and ∼47% (SUMO2) of quantified proteins were >2-fold more abundant in the purified TAP-SUMO material compared with the purified TAP-only material, and ∼36% (SUMO1) and ∼25% (SUMO2) of quantified proteins varied >2-fold in abundance in the purified TAP-SUMO material following IAV infection (Figures S3F and S3G). Together, this suggests that a large proportion of identified and quantified proteins in the purified, but not crude, samples show specific changes in abundance relating to SUMO modification status, as well as a dependence upon infection for SUMO conjugation state.

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