Signalling pathways mediating the transduction of information between cells are essential for development, cellular differentiation and homeostasis1. Their dysregulation is also frequently associated with human malignancies. The Janus tyrosine kinase/signal transducer and activator of transcription (JAK/STAT) pathway represents one such signalling cascade whose evolutionarily conserved roles include cell proliferation and haematopoiesis2. Here we describe a systematic genome-wide survey for genes required for JAK/STAT pathway activity. Analysis of 20,026 RNA interference (RNAi)-induced phenotypes in cultured Drosophila melanogaster haemocyte-like cells identified interacting genes encoding 4 known and 86 previously uncharacterized proteins. Subsequently, cell-based epistasis experiments were used to classify these proteins on the basis of their interaction with known components of the signalling cascade. In addition to multiple human disease gene homologues, we have found the tyrosine phosphatase Ptp61F and the Drosophila homologue of BRWD3, a bromo-domain-containing protein disrupted in leukaemia3. Moreover, in vivo analysis demonstrates that disrupted dBRWD3 and overexpressed Ptp61F function as suppressors of leukaemia-like blood cell tumours. This screen represents a comprehensive identification of novel loci required for JAK/STAT signalling and provides molecular insights into an important pathway relevant for human cancer. Human homologues of identified pathway modifiers may constitute targets for therapeutic interventions.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Brivanlou, A. H. & Darnell, J. E. Jr Signal transduction and the control of gene expression. Science 295, 813–818 (2002)
Hombria, J. C. & Brown, S. The fertile field of Drosophila Jak/STAT signalling. Curr. Biol. 12, R569–R575 (2002)
Kalla, C. et al. Translocation t(X;11)(q13;q23) in B-cell chronic lymphocytic leukemia disrupts two novel genes. Genes Chromosom. Cancer 42, 128–143 (2005)
Binari, R. & Perrimon, N. Stripe-specific regulation of pair-rule genes by hopscotch, a putative Jak family tyrosine kinase in Drosophila. Genes Dev. 8, 300–312 (1994)
Harrison, D. A., McCoon, P. E., Binari, R., Gilman, M. & Perrimon, N. Drosophila unpaired encodes a secreted protein that activates the JAK signalling pathway. Genes Dev. 12, 3252–3263 (1998)
Hou, X. S., Melnick, M. B. & Perrimon, N. Marelle acts downstream of the Drosophila HOP/JAK kinase and encodes a protein similar to the mammalian STATs. Cell 84, 411–419 (1996)
Lagueux, M., Perrodou, E., Levashina, E. A., Capovilla, M. & Hoffmann, J. A. Constitutive expression of a complement-like protein in toll and JAK gain-of-function mutants of Drosophila. Proc. Natl Acad. Sci. USA 97, 11427–11432 (2000)
Boutros, M., Agaisse, H. & Perrimon, N. Sequential activation of signalling pathways during innate immune responses in Drosophila. Dev. Cell 3, 711–722 (2002)
Meister, M. & Lagueux, M. Drosophila blood cells. Cell. Microbiol. 5, 573–580 (2003)
Mukherjee, T., Castelli-Gair Hombría, J. & Zeidler, M. P. Opposing roles for Drosophila JAK/STAT signalling during cellular proliferation. Oncogene 24, 2503–2511 (2005)
Brown, S., Hu, N. & Castelli-Gair Hombria, J. Identification of the first invertebrate interleukin JAK/STAT receptor, the Drosophila gene domeless. Curr. Biol. 11, 1700–1705 (2001)
Yan, R., Small, S., Desplan, C., Dearolf, C. R. & Darnell, J. E. Jr Identification of a Stat gene that functions in Drosophila development. Cell 84, 421–430 (1996)
Karsten, P., Hader, S. & Zeidler, M. Cloning and expression of Drosophila SOCS36E and its potential regulation by the JAK/STAT pathway. Mech. Dev. 117, 343–346 (2002)
Betz, A., Lampen, N., Martinek, S., Young, M. W. & Darnell, J. E. Jr A Drosophila PIAS homologue negatively regulates stat92E. Proc. Natl Acad. Sci. USA 98, 9563–9568 (2001)
Mesilaty-Gross, S., Reich, A., Motro, B. & Wides, R. The Drosophila STAM gene homolog is in a tight gene cluster, and its expression correlates to that of the adjacent gene ial. Gene 231, 173–186 (1999)
Kamath, R. S. et al. Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421, 231–237 (2003)
Boutros, M. et al. Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303, 832–835 (2004)
Kittler, R. et al. An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division. Nature 432, 1036–1040 (2004)
Hannon, G. J. & Rossi, J. J. Unlocking the potential of the human genome with RNA interference. Nature 431, 371–378 (2004)
Kwon, E. J. et al. Transcriptional regulation of the Drosophila raf proto-oncogene by Drosophila STAT during development and in immune response. J. Biol. Chem. 275, 19824–19830 (2000)
Clemens, J. C. et al. Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways. Proc. Natl Acad. Sci. USA 97, 6499–6503 (2000)
Sefton, L., Timmer, J. R., Zhang, Y., Beranger, F. & Cline, T. W. An extracellular activator of the Drosophila JAK/STAT pathway is a sex-determination signal element. Nature 405, 970–973 (2000)
Luo, H., Hanratty, W. P. & Dearolf, C. R. An amino acid substitution in the Drosophila hopTum-l Jak kinase causes leukemia-like hematopoietic defects. EMBO J. 14, 1412–1420 (1995)
Spradling, A. C. et al. The Berkeley Drosophila genome project gene disruption project: single P-element insertions mutating 25% of vital Drosophila genes. Genetics 153, 135–177 (1999)
Bach, E. A., Vincent, S., Zeidler, M. P. & Perrimon, N. A sensitized genetic screen to identify novel regulators and components of the Drosophila janus kinase/signal transducer and activator of transcription pathway. Genetics 165, 1149–1166 (2003)
Harrison, D. A., Binari, R., Nahreini, T. S., Gilman, M. & Perrimon, N. Activation of a Drosophila Janus kinase (JAK) causes hematopoietic neoplasia and developmental defects. EMBO J. 14, 2857–2865 (1995)
Yan, R., Luo, H., Darnell, J. E. Jr & Dearolf, C. R. A JAK-STAT pathway regulates wing vein formation in Drosophila. Proc. Natl Acad. Sci. USA 93, 5842–5847 (1996)
McLaughlin, S. & Dixon, J. E. Alternative splicing gives rise to a nuclear protein tyrosine phosphatase in Drosophila. J. Biol. Chem. 268, 6839–6842 (1993)
O'Shea, J. J., Pesu, M., Borie, D. C. & Changelian, P. S. A new modality for immunosuppression: targeting the JAK/STAT pathway. Nature Rev. Drug Discov. 3, 555–564 (2004)
Levy, D. E. & Darnell, J. E. Jr Stats: transcriptional control and biological impact. Nature Rev. Mol. Cell Biol. 3, 651–662 (2002)
We are grateful to R. Paro for providing reagents to establish the genome-wide RNAi library. We wish to thank M. Yamayuchi and P. Karsten for the original 2 × DrafSTAT(wt) and Socs36E promoter plasmids, C. Hunter for the sid-1 plasmid, K. Bartscherer for advice and help with tissue culture experiments, T. Horn for bioinformatics support, and M. Stricker, B. Mosterman, I. Plischke and S. Häder for technical help. We thank H. Jäckle, S. Cohen, M. Osborn, R. Paro and N. Pelte for comments on the manuscript. P.M. was supported by a fellowship from the German National Academic Foundation (Studienstiftung). Research in M.B. and M.P.Z. laboratories was supported in part by Emmy-Noether grants from the Deutsche Forschungsgemeinschaft.Author Contributions This work has been a collaborative effort between the groups of M.B. and M.P.Z.; the Boutros laboratory contributing the functional genomic expertise to dissect signalling pathways and the Zeidler laboratory expertise working with JAK/STAT signalling.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
Description of Supplementary Methods including: constructs and pathway reporter; genome-wide RNAi library; high-throughput RNAi screening; computational analysis; Sequence analysis; epistasis experiments and Ptp61F phenotype in cells; and genetics This file also contains additional references and Supplementary Figure Legends. (DOC 196 kb)
Supplementary Table S1–S7. (PDF 428 kb)
Overview of primary RNAi screen data. (PDF 1361 kb)
Loss of JAK/STAT pathway components and hopTuml induced tumour formation. (PDF 451 kb)
About this article
Cite this article
Müller, P., Kuttenkeuler, D., Gesellchen, V. et al. Identification of JAK/STAT signalling components by genome-wide RNA interference. Nature 436, 871–875 (2005). https://doi.org/10.1038/nature03869
Bromodomain and BET family proteins as epigenetic targets in cancer therapy: their degradation, present drugs, and possible PROTACs
RSC Advances (2021)
Genetic determinants of antiviral immunity in dipteran insects – Compiling the experimental evidence
Developmental & Comparative Immunology (2021)
FEBS Open Bio (2020)
Development of Multiplexed Immuno-N-Terminomics to Reveal the Landscape of Proteolytic Processing in Early Embryogenesis of Drosophila melanogaster
Analytical Chemistry (2020)
Biochemical Society Transactions (2020)