Phosphorylation of human Jak3 at tyrosines 904 and 939 positively regulates its activity.
Janus tyrosine kinase 3 (Jak3) is essential for signaling by interleukin-2 (IL-2) family cytokines and proper immune function. Dysfunctional regulation of Jak3 may result in certain disease states. However, the molecular mechanisms governing Jak3 activation are not fully understood. In this study, we used a functional-proteomics approach to identify two novel tyrosine phosphorylation sites within Jak3, Y904 and Y939, which are conserved among Jak family proteins. By using phosphospecific antibodies, both residues were observed to be rapidly induced by stimulation of cells with IL-2 or other gammac cytokines. Mechanistic studies indicated that Y904 and Y939 regulate Jak3 activities. A phenylalanine substitution at either site greatly reduced Jak3 kinase activity in vitro and its ability to phosphorylate signal transducer and activator of transcription 5 (Stat5) in vivo, suggesting that phosphorylation of these previously unrecognized residues positively regulates Jak3 activity. Y904 and Y939 were required for optimal ATP usage by Jak3, while phosphorylation of Y939 preferentially promoted Stat5 activity in intact cells. Together, these findings demonstrate positive functional roles for two novel Jak3 phosphoregulatory sites which may be similarly important for other Jak family members. Identification of these sites also provides new therapeutic opportunities to modulate Jak3 function.
Cheng
Hanyin
H
Department of Biological Sciences, Biosciences Building, University of Texas at El Paso, 500 W. University Ave., El Paso, TX 79902, USA.
Ross
Jeremy A
JA
Frost
Jeffrey A
JA
Kirken
Robert A
RA
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002460:Cell Line / Q000187:drug effects; D017124:Conserved Sequence; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D016906:Interleukin-9 / Q000494:pharmacology; D053616:Janus Kinase 3 / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000502:physiology*; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000187:drug effects / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D050799:STAT5 Transcription Factor / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D025521:Tumor Suppressor Proteins / Q000378:metabolism
C508607:IL9 protein, human; D007376:Interleukin-2; D016906:Interleukin-9; D011994:Recombinant Proteins; D050799:STAT5 Transcription Factor; C494092:STAT5A protein, human; D025521:Tumor Suppressor Proteins; D019000:Phosphotyrosine; D000255:Adenosine Triphosphate; C507933:JAK3 protein, human; D053616:Janus Kinase 3