title: Interleukin-3-induced activation of the JAK/STAT pathway is prolonged by proteasome inhibitors.
abstract: One facet of cytokine receptor signaling involves the activation of signal transducers and activators of transcription (STATs). STATs are rapidly activated via tyrosine phosphorylation by Janus kinase (JAK) family members and subsequently inactivated within a short period. We investigated the effect of proteasome inhibition on interleukin-3 (IL-3) activation of the JAK/STAT pathway following stimulation of Ba/F3 cells. Treatment of Ba/F3 cells with the proteasome inhibitor, N-acetyl-L-leucinyl-L-leucinyl-norleucinal (LLnL), led to stable tyrosine phosphorylation of the IL-3 receptor, beta common (betac), and STAT5 following stimulation. The effects of LLnL were not restricted to the JAK/STAT pathway, as Shc and mitogen-activated protein kinase (MAPK) phosphorylation were also prolonged in LLnL-treated cells. Further investigation showed these stable phosphorylation events were the result of prolonged activation of JAK2 and JAK1. These observations were confirmed using pharmacologic inhibitors. In the presence of LLnL, stable phosphorylation of STAT5 and betac was abrogated if the tyrosine kinase inhibitor, staurosporine, was added. The effect of staurosporine on STAT5 phosphorylation could be overcome if the phosphatase inhibitor, vanadate, was also added, suggesting phosphorylated STAT5 could be stabilized by phosphatase, but not by proteasome inhibition per se. These observations are consistent with the hypothesis that proteasome-mediated protein degradation can modulate the activity of the JAK/STAT pathway by regulating the deactivation of JAK.
authors.affiliation: Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
authors.forename: B A
authors.initials: BA
authors.lastname: Callus
authors.forename: B
authors.initials: B
authors.lastname: Mathey-Prevot
mesh_terms: D000818:Animals; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000502:physiology*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology*; D053614:Janus Kinase 2; D007976:Leupeptins / Q000494:pharmacology; D051379:Mice; D008894:Milk Proteins; D009097:Multienzyme Complexes / Q000502:physiology*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D046988:Proteasome Endopeptidase Complex; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D016185:Receptors, Interleukin-3 / Q000378:metabolism; D050799:STAT5 Transcription Factor; D015398:Signal Transduction; D019311:Staurosporine / Q000494:pharmacology; D015534:Trans-Activators / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology
chemical_list: D015853:Cysteine Proteinase Inhibitors; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D007377:Interleukin-3; D007976:Leupeptins; D008894:Milk Proteins; D009097:Multienzyme Complexes; D011518:Proto-Oncogene Proteins; D016185:Receptors, Interleukin-3; D050799:STAT5 Transcription Factor; D015534:Trans-Activators; D014452:Ubiquitins; C063261:acetylleucyl-leucyl-norleucinal; D014638:Vanadates; D011505:Protein-Tyrosine Kinases; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2; D010749:Phosphoprotein Phosphatases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D019311:Staurosporine