Synergistic expression of the CXCL10 gene in response to IL-1β and IFN-γ Involves NF-κB, phosphorylation of STAT1 at Tyr701, and acetylation of histones H3 and H4

Susan J. Burke, Matthew R. Goff, Danhong Lu, David Proud, Michael Karlstad, J. Jason Collier

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The CXCL10 gene encodes a peptide that chemoattracts a variety of leukocytes associated with type 1 and type 2 diabetes. The present study was undertaken to determine the molecular mechanisms required for expression of the CXCL10 gene in response to IL-1β and IFN-γ using rat islets and b cell lines. IL-1β induced the expression of the CXCL10 gene and promoter activity, whereas the combination of IL-1β plus IFN-γ was synergistic. Small interfering RNA - mediated suppression of NF-κB p65 markedly inhibited the ability of cytokines to induce the expression of the CXCL10 gene, whereas targeting STAT1 only diminished the synergy provided by IFN-γ. Furthermore, we found that a JAK1 inhibitor dose dependently reduced IFN-γ - controlled CXCL10 gene expression and promoter activity, concomitant with a decrease in STAT1 phosphorylation at Tyr701. We further discovered that, although the Tyr701 phosphorylation site is inducible (within 15 min of IFN-γ exposure), the Ser727 site within STAT1 is constitutively phosphorylated. Thus, we generated single-mutant STAT1 Y701F and double-mutant STAT1 Y701F/S727A adenoviruses. Using these recombinant adenoviruses, we determined that overexpression of either the single- or double-mutant STAT1 decreased the IFN-γ - mediated potentiation of CXCL10 gene expression, promoter activity, and secretion of protein. Moreover, the Ser727 phosphorylation was neither contingent on a functional Y701 site in β cells nor was it required for cytokine-mediated expression of the CXCL10 gene. We conclude that the synergism of IL-1β and IFN-γ to induce expression of the CXCL10 gene requires NF-κB, STAT1 phosphorylated at Tyr701, recruitment of coactivators, and acetylation of histones H3 and H4.

Original languageEnglish (US)
Pages (from-to)323-336
Number of pages14
JournalJournal of Immunology
Volume191
Issue number1
DOIs
StatePublished - Jul 1 2013

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Acetylation
Interleukin-1
Histones
Phosphorylation
Gene Expression
Adenoviridae
Cytokines
Gene Targeting
Type 1 Diabetes Mellitus
Islets of Langerhans
Type 2 Diabetes Mellitus
Small Interfering RNA
Leukocytes
Cell Line
Peptides
Genes
Proteins

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Synergistic expression of the CXCL10 gene in response to IL-1β and IFN-γ Involves NF-κB, phosphorylation of STAT1 at Tyr701, and acetylation of histones H3 and H4. / Burke, Susan J.; Goff, Matthew R.; Lu, Danhong; Proud, David; Karlstad, Michael; Collier, J. Jason.

In: Journal of Immunology, Vol. 191, No. 1, 01.07.2013, p. 323-336.

Research output: Contribution to journalArticle

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abstract = "The CXCL10 gene encodes a peptide that chemoattracts a variety of leukocytes associated with type 1 and type 2 diabetes. The present study was undertaken to determine the molecular mechanisms required for expression of the CXCL10 gene in response to IL-1β and IFN-γ using rat islets and b cell lines. IL-1β induced the expression of the CXCL10 gene and promoter activity, whereas the combination of IL-1β plus IFN-γ was synergistic. Small interfering RNA - mediated suppression of NF-κB p65 markedly inhibited the ability of cytokines to induce the expression of the CXCL10 gene, whereas targeting STAT1 only diminished the synergy provided by IFN-γ. Furthermore, we found that a JAK1 inhibitor dose dependently reduced IFN-γ - controlled CXCL10 gene expression and promoter activity, concomitant with a decrease in STAT1 phosphorylation at Tyr701. We further discovered that, although the Tyr701 phosphorylation site is inducible (within 15 min of IFN-γ exposure), the Ser727 site within STAT1 is constitutively phosphorylated. Thus, we generated single-mutant STAT1 Y701F and double-mutant STAT1 Y701F/S727A adenoviruses. Using these recombinant adenoviruses, we determined that overexpression of either the single- or double-mutant STAT1 decreased the IFN-γ - mediated potentiation of CXCL10 gene expression, promoter activity, and secretion of protein. Moreover, the Ser727 phosphorylation was neither contingent on a functional Y701 site in β cells nor was it required for cytokine-mediated expression of the CXCL10 gene. We conclude that the synergism of IL-1β and IFN-γ to induce expression of the CXCL10 gene requires NF-κB, STAT1 phosphorylated at Tyr701, recruitment of coactivators, and acetylation of histones H3 and H4.",
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