IFN-?, a cytokine produced primarily by NK cells and by Th1 CD4 and CD8 T cells, plays a central role in the immune response to infection with intracellular pathogens. Conversely, protection against extracellular metazoans is dependent on Th2 CD4 T cells and the cytokines they produce, which include IL-4, IL-13 and IL-5. Expression of IFN-? and of Th2 cytokines is primarily regulated at the level of transcription. While transcription factors that govern IFN-? expression have in recent years been elucidated in considerable detail, very little is known regarding where and how they act. By contrast to IFN-?, regulation of the Th2 cytokines IL-4, IL-13 and IL-5, which are clustered into a single ~150 kb locus in mammals, is now understood in considerable molecular detail. In this application, we propose to gain a similar level of knowledge regarding IFN-? by comprehensively identifying the regulatory elements governing IFN-? expression and the functional boundaries of the Ifng locus through 3 Aims: ? ? Aim 1: Identify regulatory elements governing the expression of IFN-? through comprehensive chromatin profiling of the murine Ifng locus. Hypothesis: DNase hypersensitive sites and regions enriched in transcriptionally favorable histone modifications extend ~50 kb upstream and downstream of the murine Ifng gene and identify regulatory elements governing IFN-? expression. ? ? Aim 2: Define the functional boundaries of the Ifng locus using BAG transgenes to complement the defect in IFN-? knockout (GKO) mice. Hypothesis: BAG transgenes containing the murine Ifng gene and extending >= 50 kb 5' and 3' will reconstitute proper IFN-? expression and thereby complement the immunological defect in GKO mice. ? ? Aim 3: Identify the mechanisms bv which the transcriptional regulatory elements within the Ifng locus influence IFN-? expression and the key transcription factors that act at these sites. Hypotheses: T-bet, STAT4, NFAT, NF-?B and AP-1 bind not only to the Ifng promoter but to additional upstream and downstream regulatory elements, thereby affecting IFN-? expression; Ifng5'CNS 2 and 3 and/or Ifng3'CNS3 help to insulate the Ifng locus from the influence of surrounding genes. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI071272-07
Application #
7247957
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Mallia, Conrad M
Project Start
2000-05-01
Project End
2011-06-30
Budget Start
2007-07-01
Budget End
2008-06-30
Support Year
7
Fiscal Year
2007
Total Cost
$336,414
Indirect Cost
Name
University of Washington
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Oestreich, Kenneth J; Mohn, Sarah E; Weinmann, Amy S (2012) Molecular mechanisms that control the expression and activity of Bcl-6 in TH1 cells to regulate flexibility with a TFH-like gene profile. Nat Immunol 13:405-11
Oestreich, Kenneth J; Weinmann, Amy S (2012) Transcriptional mechanisms that regulate T helper 1 cell differentiation. Curr Opin Immunol 24:191-5
Oestreich, Kenneth J; Huang, Albert C; Weinmann, Amy S (2011) The lineage-defining factors T-bet and Bcl-6 collaborate to regulate Th1 gene expression patterns. J Exp Med 208:1001-13
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Miller, Sara A; Mohn, Sarah E; Weinmann, Amy S (2010) Jmjd3 and UTX play a demethylase-independent role in chromatin remodeling to regulate T-box family member-dependent gene expression. Mol Cell 40:594-605
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Rowell, Emily; Merkenschlager, Matthias; Wilson, Christopher B (2008) Long-range regulation of cytokine gene expression. Curr Opin Immunol 20:272-80
Merkenschlager, Matthias; Wilson, Christopher B (2008) RNAi and chromatin in T cell development and function. Curr Opin Immunol 20:131-8

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