Proper function of cytokine signaling pathways is critical for immunoregulation, hematopoiesis, and cytokine- directed inflammation and growth. Most signals from type I and type II cytokines are mediated by the Janus kinase (Jak) family of non-receptor tyrosine kinases. Our plan is to discover the molecular basis for cytoplasmic signal transduction from the interleukin-2 family cytokines through Jak3. We will use structural biology to provide the first description of Jak interactions with cytokine receptor cytoplasmic tails, to investigate Jak kinase domain specificity, regulation and inhibition, and to describe the role of the PERM domain in kinase activation. Specifically, we will focus our work in three distinct areas.
In Aim 1 we will determine crystal structures of the kinase domain of Jak3. Modulation of the immune response by suppression of Jak3 kinase activity has clinical precedence. We will determine the mode of binding for Jak3 tyrosine kinase inhibitors and aid the discovery of further inhibitors that may be useful as immunomodulatory and anti-proliferative therapies. We will also discover the mode of Jak3 binding to a Jak3-preferred substrate and discover the inactive conformation of the kinase.
In Aim 2, we will determine the structural basis for the almost exclusive Jak3 association with the interleukin-2 receptor gamma subunit. For this we will solve crystal structures that describe the interaction of the Jak3 PERM domain with peptides from the interleukin-2 receptor gamma subunit and investigate their binding affinities. Finally, following our structural studies on the kinase and PERM domains of Jak3, in Aim 3 we will determine the structural basis for Jak3 kinase domain association with the Jak3 FERM domain. We will also test the role of this interface as a suggested Jak3 active state stabilizing mechanism. These studies will directly impact healthcare stimulating the development of improved immunomodulatory and anti-proliferative therapies and by enhancing molecular-level understanding of cytokine signaling pathways.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI075133-05
Application #
8080988
Study Section
Cellular and Molecular Immunology - A Study Section (CMIA)
Program Officer
Mallia, Conrad M
Project Start
2007-06-01
Project End
2013-05-31
Budget Start
2011-06-01
Budget End
2013-05-31
Support Year
5
Fiscal Year
2011
Total Cost
$430,831
Indirect Cost
Name
Yale University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Gifford, Stacey M; Liu, Weizhi; Mader, Christopher C et al. (2014) Two amino acid residues confer different binding affinities of Abelson family kinase SRC homology 2 domains for phosphorylated cortactin. J Biol Chem 289:19704-13
Deng, Yang; Alicea-Velázquez, Nilda L; Bannwarth, Ludovic et al. (2014) Global analysis of human nonreceptor tyrosine kinase specificity using high-density peptide microarrays. J Proteome Res 13:4339-46
Alicea-Velazquez, Nilda L; Jakoncic, Jean; Boggon, Titus J (2013) Structure-guided studies of the SHP-1/JAK1 interaction provide new insights into phosphatase catalytic domain substrate recognition. J Struct Biol 181:243-51
Alicea-Velazquez, Nilda L; Boggon, Titus J (2013) SHP family protein tyrosine phosphatases adopt canonical active-site conformations in the apo and phosphate-bound states. Protein Pept Lett 20:1039-48
Li, Xiaofeng; Ji, Weidong; Zhang, Rong et al. (2011) Molecular recognition of leucine-aspartate repeat (LD) motifs by the focal adhesion targeting homology domain of cerebral cavernous malformation 3 (CCM3). J Biol Chem 286:26138-47
Alicea-Velazquez, Nilda L; Boggon, Titus J (2011) The use of structural biology in Janus kinase targeted drug discovery. Curr Drug Targets 12:546-55
Zhang, Zhenfeng; Stiegler, Amy L; Boggon, Titus J et al. (2010) EGFR-mutated lung cancer: a paradigm of molecular oncology. Oncotarget 1:497-514
Bae, Jae Hyun; Boggon, Titus J; Tomé, Francisco et al. (2010) Asymmetric receptor contact is required for tyrosine autophosphorylation of fibroblast growth factor receptor in living cells. Proc Natl Acad Sci U S A 107:2866-71
Cornejo, Melanie G; Boggon, Titus J; Mercher, Thomas (2009) JAK3: a two-faced player in hematological disorders. Int J Biochem Cell Biol 41:2376-9
Kumar, Amit; Petri, Edward T; Halmos, Balazs et al. (2008) Structure and clinical relevance of the epidermal growth factor receptor in human cancer. J Clin Oncol 26:1742-51