Janus kinases (Jaks), members of the non-receptor protein tyrosine kinase family, are key components of signaling pathways in cells of the immune system and in hematopoietic cells. Jaks are associated with the cytoplasmic domains of cytokine receptors and, upon cytokine-mediated receptor dimerization, undergo transautophosphorylation on tyrosine residues, which stimulates their tyrosine kinase activity. Activated Jaks phosphorylate STATs (signal transducers and activators of transcription), which translocate to the nucleus and serve as transcriptional activators. There are four mammalian members of the Jak family (Jak1-3 and Tyk2) which possess four domains in common: an N-terminal FERM domain, an SH2-like domain, a pseudokinase domain, and a tyrosine kinase domain. Extensive biochemical data, as well as gain-of-function mutations that cause myeloproliferative diseases/cancers, have implicated the pseudokinase domain of Jaks as crucial for maintaining a low basal level of tyrosine kinase activity. The goal of this proposal is to understand the structural/molecular mechanisms by which the pseudokinase domain negatively regulates the tyrosine kinase activity of Jak2. To achieve this goal, x-ray crystallography will be employed to determine the three-dimensional structures of the pseudokinase domain and the tandem pseudokinase and tyrosine kinase domains.
|Silvennoinen, Olli; Hubbard, Stevan R (2015) Molecular insights into regulation of JAK2 in myeloproliferative neoplasms. Blood 125:3388-92|
|Hammarén, Henrik M; Ungureanu, Daniela; Grisouard, Jean et al. (2015) ATP binding to the pseudokinase domain of JAK2 is critical for pathogenic activation. Proc Natl Acad Sci U S A 112:4642-7|
|Shan, Yibing; Gnanasambandan, Kavitha; Ungureanu, Daniela et al. (2014) Molecular basis for pseudokinase-dependent autoinhibition of JAK2 tyrosine kinase. Nat Struct Mol Biol 21:579-84|
|Silvennoinen, Olli; Ungureanu, Daniela; Niranjan, Yashavanthi et al. (2013) New insights into the structure and function of the pseudokinase domain in JAK2. Biochem Soc Trans 41:1002-7|
|Bandaranayake, Rajintha M; Ungureanu, Daniela; Shan, Yibing et al. (2012) Crystal structures of the JAK2 pseudokinase domain and the pathogenic mutant V617F. Nat Struct Mol Biol 19:754-9|