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Recruitment of histone deacetylase 3 to the interferon-A gene promoters attenuates interferon expression.

Génin P, Lin R, Hiscott J, Civas A - PLoS ONE (2012)

Bottom Line: Analysis of chromatin-protein association by Chip-QPCR demonstrated that recruitment of interferon regulatory factor (IRF)3 and IRF7, as well as TBP correlated with enhanced histone H3K9 and H3K14 acetylation, whereas recruitment of HDAC3 correlated with inhibition of histone H3K9/K14 acetylation, removal of IRF7 and TATA-binding protein (TBP) from IFN-A promoters and inhibition of virus-induced IFN-A gene transcription.Additionally, HDAC3 overexpression reduced, and HDAC3 depletion by siRNA enhanced IFN-A gene expression.Furthermore, activation of IRF7 enhanced histone H3K9/K14 acetylation and IFN-A gene expression, whereas activation of both IRF7 and IRF3 led to recruitment of HDAC3 to the IFN-A gene promoters, resulting in impaired histone H3K9 acetylation and attenuation of IFN-A gene transcription.

View Article: PubMed Central - PubMed

Affiliation: Centre National de la Recherche Scientifique-FRE3235, Paris Descartes University, Paris, France.

ABSTRACT

Background: Induction of Type I Interferon (IFN) genes constitutes an essential step leading to innate immune responses during virus infection. Sendai virus (SeV) infection of B lymphoid Namalwa cells transiently induces the transcriptional expression of multiple IFN-A genes. Although transcriptional activation of IFN-A genes has been extensively studied, the mechanism responsible for the attenuation of their expression remains to be determined.

Principal findings: In this study, we demonstrate that virus infection of Namalwa cells induces transient recruitment of HDAC3 (histone deacetylase 3) to IFN-A promoters. Analysis of chromatin-protein association by Chip-QPCR demonstrated that recruitment of interferon regulatory factor (IRF)3 and IRF7, as well as TBP correlated with enhanced histone H3K9 and H3K14 acetylation, whereas recruitment of HDAC3 correlated with inhibition of histone H3K9/K14 acetylation, removal of IRF7 and TATA-binding protein (TBP) from IFN-A promoters and inhibition of virus-induced IFN-A gene transcription. Additionally, HDAC3 overexpression reduced, and HDAC3 depletion by siRNA enhanced IFN-A gene expression. Furthermore, activation of IRF7 enhanced histone H3K9/K14 acetylation and IFN-A gene expression, whereas activation of both IRF7 and IRF3 led to recruitment of HDAC3 to the IFN-A gene promoters, resulting in impaired histone H3K9 acetylation and attenuation of IFN-A gene transcription.

Conclusion: Altogether these data indicate that reversal of histone H3K9/K14 acetylation by HDAC3 is required for attenuation of IFN-A gene transcription during viral infection.

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Expression pattern of IFN-A genes in Namalwa B cells infected by Sendai virus.(A) Type I IFN locus, delimited to 400 kbps on human chromosome 9, consists of thirteen intronless IFN-A genes located in cluster with the IFN-B and IFN-E genes (data retrieved from NCBI library, version 37.1). IFN-A genes analyzed in this study are indicated in bold in the diagram. IFN-A1 and IFN-A8 are in reverse orientation compared to other IFN-A genes. (B) IFN-A gene expression was determined by quantitative RT-QPCR analysis in Namalwa B cells mock-infected (0 h) or infected by Sendai virus for 4–16 h. IFN-A1 expression levels presented in this study represent the total amounts of both IFN-A1 and IFN-A13 mRNAs; the high percentage of sequence homology of IFN-A1 and IFN-A13 genes (99.7% in the coding region) did not allow distinction between these genes. Relative expression (RE) normalized to GAPDH mRNA levels is indicated with standard deviation values calculated in two independent experiments performed in duplicate.
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pone-0038336-g001: Expression pattern of IFN-A genes in Namalwa B cells infected by Sendai virus.(A) Type I IFN locus, delimited to 400 kbps on human chromosome 9, consists of thirteen intronless IFN-A genes located in cluster with the IFN-B and IFN-E genes (data retrieved from NCBI library, version 37.1). IFN-A genes analyzed in this study are indicated in bold in the diagram. IFN-A1 and IFN-A8 are in reverse orientation compared to other IFN-A genes. (B) IFN-A gene expression was determined by quantitative RT-QPCR analysis in Namalwa B cells mock-infected (0 h) or infected by Sendai virus for 4–16 h. IFN-A1 expression levels presented in this study represent the total amounts of both IFN-A1 and IFN-A13 mRNAs; the high percentage of sequence homology of IFN-A1 and IFN-A13 genes (99.7% in the coding region) did not allow distinction between these genes. Relative expression (RE) normalized to GAPDH mRNA levels is indicated with standard deviation values calculated in two independent experiments performed in duplicate.

Mentions: In the present study, Namalwa B lymphoid cells infected with Sendai virus (SeV) were used as a model to determine the kinetics of IFN-A gene activation, with each functional IFN-A gene present in the type I IFN locus monitored by RT-QPCR. IFN-A genes exhibited transient expression with mRNA levels accumulating between 4 and 8 h (IFN-A1, A2, A8, A10, A14, A16 and A17) or between 6 and 12 h (IFN-A5 and A7) post-infection (p.i), whereas IFN-A4, A6 and A21 genes were weakly induced (Fig. 1A-B). IFN-A gene expression decreased after 8 h and 12 h of infection, for the first and second groups, respectively. Decrease in mRNA levels was independent of actinomycin D addition, as shown for IFN-A1, A2 and A14 (Fig. S1), suggesting that attenuation of IFN-A gene expression was due to transcriptional inhibition.


Recruitment of histone deacetylase 3 to the interferon-A gene promoters attenuates interferon expression.

Génin P, Lin R, Hiscott J, Civas A - PLoS ONE (2012)

Expression pattern of IFN-A genes in Namalwa B cells infected by Sendai virus.(A) Type I IFN locus, delimited to 400 kbps on human chromosome 9, consists of thirteen intronless IFN-A genes located in cluster with the IFN-B and IFN-E genes (data retrieved from NCBI library, version 37.1). IFN-A genes analyzed in this study are indicated in bold in the diagram. IFN-A1 and IFN-A8 are in reverse orientation compared to other IFN-A genes. (B) IFN-A gene expression was determined by quantitative RT-QPCR analysis in Namalwa B cells mock-infected (0 h) or infected by Sendai virus for 4–16 h. IFN-A1 expression levels presented in this study represent the total amounts of both IFN-A1 and IFN-A13 mRNAs; the high percentage of sequence homology of IFN-A1 and IFN-A13 genes (99.7% in the coding region) did not allow distinction between these genes. Relative expression (RE) normalized to GAPDH mRNA levels is indicated with standard deviation values calculated in two independent experiments performed in duplicate.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3369917&req=5

pone-0038336-g001: Expression pattern of IFN-A genes in Namalwa B cells infected by Sendai virus.(A) Type I IFN locus, delimited to 400 kbps on human chromosome 9, consists of thirteen intronless IFN-A genes located in cluster with the IFN-B and IFN-E genes (data retrieved from NCBI library, version 37.1). IFN-A genes analyzed in this study are indicated in bold in the diagram. IFN-A1 and IFN-A8 are in reverse orientation compared to other IFN-A genes. (B) IFN-A gene expression was determined by quantitative RT-QPCR analysis in Namalwa B cells mock-infected (0 h) or infected by Sendai virus for 4–16 h. IFN-A1 expression levels presented in this study represent the total amounts of both IFN-A1 and IFN-A13 mRNAs; the high percentage of sequence homology of IFN-A1 and IFN-A13 genes (99.7% in the coding region) did not allow distinction between these genes. Relative expression (RE) normalized to GAPDH mRNA levels is indicated with standard deviation values calculated in two independent experiments performed in duplicate.
Mentions: In the present study, Namalwa B lymphoid cells infected with Sendai virus (SeV) were used as a model to determine the kinetics of IFN-A gene activation, with each functional IFN-A gene present in the type I IFN locus monitored by RT-QPCR. IFN-A genes exhibited transient expression with mRNA levels accumulating between 4 and 8 h (IFN-A1, A2, A8, A10, A14, A16 and A17) or between 6 and 12 h (IFN-A5 and A7) post-infection (p.i), whereas IFN-A4, A6 and A21 genes were weakly induced (Fig. 1A-B). IFN-A gene expression decreased after 8 h and 12 h of infection, for the first and second groups, respectively. Decrease in mRNA levels was independent of actinomycin D addition, as shown for IFN-A1, A2 and A14 (Fig. S1), suggesting that attenuation of IFN-A gene expression was due to transcriptional inhibition.

Bottom Line: Analysis of chromatin-protein association by Chip-QPCR demonstrated that recruitment of interferon regulatory factor (IRF)3 and IRF7, as well as TBP correlated with enhanced histone H3K9 and H3K14 acetylation, whereas recruitment of HDAC3 correlated with inhibition of histone H3K9/K14 acetylation, removal of IRF7 and TATA-binding protein (TBP) from IFN-A promoters and inhibition of virus-induced IFN-A gene transcription.Additionally, HDAC3 overexpression reduced, and HDAC3 depletion by siRNA enhanced IFN-A gene expression.Furthermore, activation of IRF7 enhanced histone H3K9/K14 acetylation and IFN-A gene expression, whereas activation of both IRF7 and IRF3 led to recruitment of HDAC3 to the IFN-A gene promoters, resulting in impaired histone H3K9 acetylation and attenuation of IFN-A gene transcription.

View Article: PubMed Central - PubMed

Affiliation: Centre National de la Recherche Scientifique-FRE3235, Paris Descartes University, Paris, France.

ABSTRACT

Background: Induction of Type I Interferon (IFN) genes constitutes an essential step leading to innate immune responses during virus infection. Sendai virus (SeV) infection of B lymphoid Namalwa cells transiently induces the transcriptional expression of multiple IFN-A genes. Although transcriptional activation of IFN-A genes has been extensively studied, the mechanism responsible for the attenuation of their expression remains to be determined.

Principal findings: In this study, we demonstrate that virus infection of Namalwa cells induces transient recruitment of HDAC3 (histone deacetylase 3) to IFN-A promoters. Analysis of chromatin-protein association by Chip-QPCR demonstrated that recruitment of interferon regulatory factor (IRF)3 and IRF7, as well as TBP correlated with enhanced histone H3K9 and H3K14 acetylation, whereas recruitment of HDAC3 correlated with inhibition of histone H3K9/K14 acetylation, removal of IRF7 and TATA-binding protein (TBP) from IFN-A promoters and inhibition of virus-induced IFN-A gene transcription. Additionally, HDAC3 overexpression reduced, and HDAC3 depletion by siRNA enhanced IFN-A gene expression. Furthermore, activation of IRF7 enhanced histone H3K9/K14 acetylation and IFN-A gene expression, whereas activation of both IRF7 and IRF3 led to recruitment of HDAC3 to the IFN-A gene promoters, resulting in impaired histone H3K9 acetylation and attenuation of IFN-A gene transcription.

Conclusion: Altogether these data indicate that reversal of histone H3K9/K14 acetylation by HDAC3 is required for attenuation of IFN-A gene transcription during viral infection.

Show MeSH
Related in: MedlinePlus