title: Acute lymphoblastic leukemia-associated JAK1 mutants activate the Janus kinase/STAT pathway via interleukin-9 receptor alpha homodimers.
abstract: Activating mutations in JAK1 have been reported in acute lymphoblastic leukemias, but little is known about the mechanisms involved in their constitutive activation. Here, we studied the ability of JAK1 V658F and A634D to activate the Janus kinase (JAK)/STAT pathway upon ectopic expression in HEK293 cells alone or together with the other components of the interleukin-9 receptor complex (IL-9Ralpha, gammac, and JAK3). Expression of JAK1 mutants alone failed to trigger STAT activation, but co-expression of the IL-9Ralpha chain promoted JAK1 mutant phosphorylation and STAT activation. Mutation of the FERM domain of JAK1, which is critical for cytokine receptor association, or of the single tyrosine of IL-9Ralpha involved in STAT recruitment abolished this activity, indicating that JAK1 mutants need to associate with a functional IL-9Ralpha to activate STAT factors. Several lines of evidence indicated that IL-9Ralpha homodimerization was involved in this process. IL-9Ralpha variants with mutations of the JAK-interacting BOX1 region not only failed to promote JAK1 activation but also acted as dominant negative forms reverting the effect of wild-type IL-9Ralpha. Coimmunoprecipitation experiments also showed the formation of IL-9Ralpha homodimers. Interestingly, STAT activation was partially inhibited by expression of gammac, suggesting that overlapping residues are involved in IL-9Ralpha homodimerization and IL-9Ralpha/gammac heterodimerization. Co-expression of wild-type JAK3 partially reverted the inhibition by gammac, indicating that JAK3 cooperates with JAK1 mutants within the IL-9 receptor complex. Similar results were observed with IL-2Rbeta. Taken together, our results show that IL-9Ralpha and IL-2Rbeta homodimers efficiently mediate constitutive activation of ALL-associated JAK1 mutants.
authors.affiliation: Ludwig Institute for Cancer Research, Brussels Branch and de Duve Institute, Université Catholique de Louvain, B-1200 Brussels, Belgium.
authors.forename: Tekla
authors.initials: T
authors.lastname: Hornakova
authors.forename: Judith
authors.initials: J
authors.lastname: Staerk
authors.forename: Yohan
authors.initials: Y
authors.lastname: Royer
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Flex
authors.forename: Marco
authors.initials: M
authors.lastname: Tartaglia
authors.forename: Stefan N
authors.initials: SN
authors.lastname: Constantinescu
authors.forename: Laurent
authors.initials: L
authors.lastname: Knoops
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Renauld
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D045744:Cell Line, Tumor; D019281:Dimerization; D004789:Enzyme Activation / Q000235:genetics; D006801:Humans; D053646:Interleukin-2 Receptor beta Subunit / Q000235:genetics / Q000378:metabolism; D053613:Janus Kinase 1 / Q000235:genetics / Q000378:metabolism*; D053616:Janus Kinase 3 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D020125:Mutation, Missense; D010766:Phosphorylation / Q000235:genetics; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D053651:Receptors, Interleukin-9 / Q000235:genetics / Q000378:metabolism*; D050791:STAT Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C508020:IL2RB protein, human; C508045:IL9R protein, human; C508022:Il2rb protein, mouse; C508448:Il9r protein, mouse; D053646:Interleukin-2 Receptor beta Subunit; D053651:Receptors, Interleukin-9; D050791:STAT Transcription Factors; C507904:JAK1 protein, human; C507933:JAK3 protein, human; C507905:Jak1 protein, mouse; C507935:Jak3 protein, mouse; D053613:Janus Kinase 1; D053616:Janus Kinase 3