Limits...
DEAF1 is a Pellino1-interacting protein required for interferon production by Sendai virus and double-stranded RNA.

Ordureau A, Enesa K, Nanda S, Le Francois B, Peggie M, Prescott A, Albert PR, Cohen P - J. Biol. Chem. (2013)

Bottom Line: The interaction is independent of the E3 ligase activity of Pellino1, but weakened by the phosphorylation of Pellino1.DEAF1 is also needed for TLR3-dependent IFNβ production.Taken together, our results identify DEAF1 as a novel component of the signal transduction network by which dsRNA of viral origin stimulates IFNβ production.

View Article: PubMed Central - PubMed

Affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom.

ABSTRACT
Double-stranded (ds) RNA of viral origin, a ligand for Melanoma Differentiation-associated gene 5 (MDA5) and Toll-Like Receptor 3 (TLR3), induces the TANK-Binding Kinase 1 (TBK1)-dependent phosphorylation and activation of Interferon Regulatory Factor 3 (IRF3) and the E3 ubiquitin ligase Pellino1, which are required for interferon β (IFNβ) gene transcription. Here, we report that Pellino1 interacts with the transcription factor Deformed Epidermal Autoregulatory Factor 1 (DEAF1). The interaction is independent of the E3 ligase activity of Pellino1, but weakened by the phosphorylation of Pellino1. We show that DEAF1 binds to the IFNβ promoter and to IRF3 and IRF7, that it is required for the transcription of the IFNβ gene and IFNβ secretion in MEFs infected with Sendai virus or transfected with poly(I:C). DEAF1 is also needed for TLR3-dependent IFNβ production. Taken together, our results identify DEAF1 as a novel component of the signal transduction network by which dsRNA of viral origin stimulates IFNβ production.

Show MeSH

Related in: MedlinePlus

The poly(I:C)-stimulated production of IFNβ is impaired in BMDM from DEAF1−/− mice.A and B, four wild type mice and three DEAF1−/− mice were used at each time point. BMDM from wild type (WT) (black bars) or DEAF1−/− mice (white bars) were stimulated for the times indicated with poly(I:C) (10 μg/ml) and, at each time point, the total RNA was extracted from the macrophages and the mRNA encoding IFNβ (A) and CXCL10 (B) was measured by qRT-PCR. The results show the fold increase in mRNA relative to the values measured in unstimulated, wild-type BMDMs (± S.D. for triplicate determinations). C, as in A and B except that, at each time point, the amount of IFNβ secreted into the cell culture medium from wild type BMDM (black circles) or DEAF1−/− BMDM (white circles) was measured by ELISA. D, same as C, except that BMDM from wild type (WT) mice (black squares) or Pellino1[F397A] mice (white squares) (9) were used.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3750155&req=5

Figure 9: The poly(I:C)-stimulated production of IFNβ is impaired in BMDM from DEAF1−/− mice.A and B, four wild type mice and three DEAF1−/− mice were used at each time point. BMDM from wild type (WT) (black bars) or DEAF1−/− mice (white bars) were stimulated for the times indicated with poly(I:C) (10 μg/ml) and, at each time point, the total RNA was extracted from the macrophages and the mRNA encoding IFNβ (A) and CXCL10 (B) was measured by qRT-PCR. The results show the fold increase in mRNA relative to the values measured in unstimulated, wild-type BMDMs (± S.D. for triplicate determinations). C, as in A and B except that, at each time point, the amount of IFNβ secreted into the cell culture medium from wild type BMDM (black circles) or DEAF1−/− BMDM (white circles) was measured by ELISA. D, same as C, except that BMDM from wild type (WT) mice (black squares) or Pellino1[F397A] mice (white squares) (9) were used.

Mentions: DEAF1−/− mice are rarely born alive, and the few survivors are very small. Nevertheless, a few experiments with BMDM from DEAF1−/− mice were performed, which showed that the poly(I:C)-stimulated production of IFNβ mRNA (Fig. 9A) or CXCL10 (Fig. 9B) was reduced considerably compared with BMDM from wild-type mice. The reduction in IFNβ secretion (Fig. 9C) was similar to that observed in BMDM from mice expressing the E3 ligase-inactive Pellino1[F397A] mutant (Fig. 9D) (9).


DEAF1 is a Pellino1-interacting protein required for interferon production by Sendai virus and double-stranded RNA.

Ordureau A, Enesa K, Nanda S, Le Francois B, Peggie M, Prescott A, Albert PR, Cohen P - J. Biol. Chem. (2013)

The poly(I:C)-stimulated production of IFNβ is impaired in BMDM from DEAF1−/− mice.A and B, four wild type mice and three DEAF1−/− mice were used at each time point. BMDM from wild type (WT) (black bars) or DEAF1−/− mice (white bars) were stimulated for the times indicated with poly(I:C) (10 μg/ml) and, at each time point, the total RNA was extracted from the macrophages and the mRNA encoding IFNβ (A) and CXCL10 (B) was measured by qRT-PCR. The results show the fold increase in mRNA relative to the values measured in unstimulated, wild-type BMDMs (± S.D. for triplicate determinations). C, as in A and B except that, at each time point, the amount of IFNβ secreted into the cell culture medium from wild type BMDM (black circles) or DEAF1−/− BMDM (white circles) was measured by ELISA. D, same as C, except that BMDM from wild type (WT) mice (black squares) or Pellino1[F397A] mice (white squares) (9) were used.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750155&req=5

Figure 9: The poly(I:C)-stimulated production of IFNβ is impaired in BMDM from DEAF1−/− mice.A and B, four wild type mice and three DEAF1−/− mice were used at each time point. BMDM from wild type (WT) (black bars) or DEAF1−/− mice (white bars) were stimulated for the times indicated with poly(I:C) (10 μg/ml) and, at each time point, the total RNA was extracted from the macrophages and the mRNA encoding IFNβ (A) and CXCL10 (B) was measured by qRT-PCR. The results show the fold increase in mRNA relative to the values measured in unstimulated, wild-type BMDMs (± S.D. for triplicate determinations). C, as in A and B except that, at each time point, the amount of IFNβ secreted into the cell culture medium from wild type BMDM (black circles) or DEAF1−/− BMDM (white circles) was measured by ELISA. D, same as C, except that BMDM from wild type (WT) mice (black squares) or Pellino1[F397A] mice (white squares) (9) were used.
Mentions: DEAF1−/− mice are rarely born alive, and the few survivors are very small. Nevertheless, a few experiments with BMDM from DEAF1−/− mice were performed, which showed that the poly(I:C)-stimulated production of IFNβ mRNA (Fig. 9A) or CXCL10 (Fig. 9B) was reduced considerably compared with BMDM from wild-type mice. The reduction in IFNβ secretion (Fig. 9C) was similar to that observed in BMDM from mice expressing the E3 ligase-inactive Pellino1[F397A] mutant (Fig. 9D) (9).

Bottom Line: The interaction is independent of the E3 ligase activity of Pellino1, but weakened by the phosphorylation of Pellino1.DEAF1 is also needed for TLR3-dependent IFNβ production.Taken together, our results identify DEAF1 as a novel component of the signal transduction network by which dsRNA of viral origin stimulates IFNβ production.

View Article: PubMed Central - PubMed

Affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom.

ABSTRACT
Double-stranded (ds) RNA of viral origin, a ligand for Melanoma Differentiation-associated gene 5 (MDA5) and Toll-Like Receptor 3 (TLR3), induces the TANK-Binding Kinase 1 (TBK1)-dependent phosphorylation and activation of Interferon Regulatory Factor 3 (IRF3) and the E3 ubiquitin ligase Pellino1, which are required for interferon β (IFNβ) gene transcription. Here, we report that Pellino1 interacts with the transcription factor Deformed Epidermal Autoregulatory Factor 1 (DEAF1). The interaction is independent of the E3 ligase activity of Pellino1, but weakened by the phosphorylation of Pellino1. We show that DEAF1 binds to the IFNβ promoter and to IRF3 and IRF7, that it is required for the transcription of the IFNβ gene and IFNβ secretion in MEFs infected with Sendai virus or transfected with poly(I:C). DEAF1 is also needed for TLR3-dependent IFNβ production. Taken together, our results identify DEAF1 as a novel component of the signal transduction network by which dsRNA of viral origin stimulates IFNβ production.

Show MeSH
Related in: MedlinePlus