Interferon gamma (IFN-gamma) plays a crucial role in regulation of the immune system and is active on virtually all tissues. Its actions are mediated by its receptor that is the subject of this application. We previously identified two chains (IFN-gammaR1 and IFN-gammaR2) that are required for receptor activity. The current proposal focuses on the structure of the chains of the IFN-gamma receptor complex in live cells in the presence and absence of ligand.
The specific aims directed to these goals are: to determine if all the IFN-gamma, receptor chains preassociated; the role of the Jak kinases in preassembly; determination of the sites on the receptor chains for preassembly; role of Stat1 in movement of the intracellular domains; determination of the kinetics of change in receptor structure after ligand binding; analysis of the kinetics and interactions of the downstream signaling components with the receptor complex; determine the sites of the receptor that serve as sites for cross-talk between the IFN-gamma and IFN-alpha receptor complexes. The configuration of the receptor chains will be studied by fluorescence resonance energy transfer in live cells. Because IFN-gamma plays a critical role in cellular and immune function, understanding all these aspects of its action will lead to better strategies to attack a variety of diseases such as cancers, and infectious and autoimmune diseases. Further application of this technology should lead to new rapid methods for high throughput screening and for identifying protein-protein interactions in live cells in real-time.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI059465-01
Application #
6758085
Study Section
Allergy and Immunology Study Section (ALY)
Program Officer
Rathbun, Gary
Project Start
2004-04-01
Project End
2008-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
1
Fiscal Year
2004
Total Cost
$349,875
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Genetics
Type
Schools of Medicine
DUNS #
617022384
City
Piscataway
State
NJ
Country
United States
Zip Code
08854
Krause, Christopher D; Izotova, Lara S; Pestka, Sidney (2013) Analytical use of multi-protein Fluorescence Resonance Energy Transfer to demonstrate membrane-facilitated interactions within cytokine receptor complexes. Cytokine 64:298-309
Krause, Christopher D; Digioia, Gina; Izotova, Lara S et al. (2013) Improving the spectral analysis of Fluorescence Resonance Energy Transfer in live cells: application to interferon receptors and Janus kinases. Cytokine 64:272-85
Krause, Christopher D; Digioia, Gina; Izotova, Lara S et al. (2013) Ligand-independent interaction of the type I interferon receptor complex is necessary to observe its biological activity. Cytokine 64:286-97
Sarkar, Srijata; Han, Junfeng; Sinsimer, Kristina S et al. (2011) RNA-binding protein AUF1 regulates lipopolysaccharide-induced IL10 expression by activating IkappaB kinase complex in monocytes. Mol Cell Biol 31:602-15
Knapinska, Anna M; Gratacós, Frances M; Krause, Christopher D et al. (2011) Chaperone Hsp27 modulates AUF1 proteolysis and AU-rich element-mediated mRNA degradation. Mol Cell Biol 31:1419-31
Sarkar, Srijata; Sinsimer, Kristina S; Foster, Randi L et al. (2008) AUF1 isoform-specific regulation of anti-inflammatory IL10 expression in monocytes. J Interferon Cytokine Res 28:679-91
Sinsimer, Kristina S; Gratacos, Frances M; Knapinska, Anna M et al. (2008) Chaperone Hsp27, a novel subunit of AUF1 protein complexes, functions in AU-rich element-mediated mRNA decay. Mol Cell Biol 28:5223-37
Krause, Christopher D; Mei, Erwen; Mirochnitchenko, Olga et al. (2006) Interactions among the components of the interleukin-10 receptor complex. Biochem Biophys Res Commun 340:377-85
Krause, Christopher D; Lavnikova, Natasha; Xie, Junxia et al. (2006) Preassembly and ligand-induced restructuring of the chains of the IFN-gamma receptor complex: the roles of Jak kinases, Stat1 and the receptor chains. Cell Res 16:55-69
Krause, Christopher D; He, Wen; Kotenko, Sergei et al. (2006) Modulation of the activation of Stat1 by the interferon-gamma receptor complex. Cell Res 16:113-23

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