Multiple kinases in the interferon-gamma response.
Janus kinases (JAKs) and signal transducers and activators of transcription (STATs) are essential for responses to interferons (IFNs), most cytokines, and some growth factors. JAK/STAT signaling is not, however, sufficient for a full IFN-gamma response. Here, a convenient, robust, and quantitative flow cytometry-based kinome-wide siRNA screen has identified nine additional kinases as required for the IFN-gamma class II HLA response, seven for an antiviral response, and two for the cytopathic response to encephalomyocarditis virus (EMCV). As one example, inhibition of the IFN-gamma response by siRNA to ataxia telangiectasia-mutated (ATM) differentially affects a spectrum of IFN-gamma-stimulated mRNAs, with inhibitions being seen as early as 1 h after IFN-gamma stimulation. The implication of ATM, with its previously recognized function in chromatin decondensation, in the control of transcription early in the IFN-gamma response highlights both a role for ATM in cytokine responses and a possible correlation with the chromatin decondensation recently observed in response to IFN-gamma in mammalian cells. This work has, therefore, revealed the simplicity, power, and convenience of quantitative flow cytometry-based siRNA screens, a requirement for ATM and multiple additional kinases in the IFN-gamma response and a possible requirement for two of these kinases in the cytopathic response to EMCV.
Watling
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Department of Oncology, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0NN, United Kingdom.
Carmo
C R
CR
Kerr
I M
IM
Costa-Pereira
A P
AP
D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins; D018797:Cell Cycle Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology*; D051379:Mice; D009369:Neoplasms / Q000201:enzymology / Q000276:immunology; D010770:Phosphotransferases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D050794:STAT1 Transcription Factor / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics; D014780:Viruses / Q000276:immunology
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D000949:Histocompatibility Antigens Class II; D034741:RNA, Small Interfering; D050794:STAT1 Transcription Factor; D025521:Tumor Suppressor Proteins; D007371:Interferon-gamma; D010770:Phosphotransferases; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C576325:Atm protein, mouse; D017346:Protein Serine-Threonine Kinases