Abstract
Signal transducers and activators of transcription (STAT) proteins are transcription factors activated by phosphorylation on tyrosine residues after cytokine stimulation. In erythropoietin receptor (EPOR)-mediated signaling, STAT5 is tyrosine-phosphorylated by EPO stimulation. Although Janus Kinase 2 (JAK2) is reported to play a crucial role in EPO-induced activation of STAT5, it is unclear whether JAK2 alone can tyrosine-phosphorylate STAT5 after EPO stimulation. Several studies indicate that STAT activation is caused by members of other families of protein tyrosine kinases such as the Src family. We previously reported that reduction of Src by induction of antisense src RNA expression suppressed EPO-promoted erythroid differentiation in K562 cells. In the present study, we explored the function of Src downstream of the EPOR-initiated signaling. Reduction of Src diminished tyrosine phosphorylation of STAT5 in K562 cells regardless of EPO treatment. The tyrosine phosphorylation level of STAT5 induced by EPO in F-36P cells was reduced in the presence of PP1 or PP2 selective Src inhibitor. In addition, the expression of dominant negative Src in F-36P cells reduced the tyrosine phosphorylation of STAT5. When Src and STAT5 were co-expressed in COS7 cells, tyrosine phosphorylation of STAT5 was observed, and tyrosine residue 694 (Tyr 694) of STAT5A was identified as the major phosphorylation site by Src. In vitro kinase assay revealed that GST-STAT5 fusion protein with the conserved C-terminal, but not the C-terminal-truncated mutant which lacks Tyr 694, was tyrosine-phosphorylated by Src. Src can thus directly tyrosine-phosphorylate the activation site of STAT5 (Tyr 694 in STAT5A), and Src may contribute to EPO-induced signal transduction via STAT5.
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References
Cao X, Tay A, Guy GR, Tan YH . 1996 Mol. Cell. Biol. 16: 1595–1603
Carlesso N, Frank DA, Griffin JD . 1996 J. Exp. Med. 183: 811–820
Chaturvedi P, Sharma S, Reddy EP . 1997 Mol. Cell. Biol. 17: 3295–3304
Chaturvedi P, Reddy MV, Reddy EP . 1998 Oncogene 16: 1749–1758
Chiba S, Takaku F, Tange T, Shibuya K, Misawa C, Sasaki K, Miyagawa K, Yazaki Y, Hirai H . 1991 Blood 78: 2261–2268
Chin H, Arai A, Wakao H, Kamiyama R, Miyasaka N, Miura O . 1998 Blood 91: 3734–3745
Coffer PJ, Koenderman L, de Groot RP . 2000 Oncogene 19: 2511–2522
Constantinescu SN, Ghaffari S, Lodish HF . 1999 Trends Endocrinol. Metab. 10: 18–23
Darnell Jr JE, Kerr IM, Stark GR . 1994 Science 264: 1415–1421
Danial NN, Pernis A, Rothman PB . 1995 Science 269: 1875–1877
de Groot RP, Raaijmakers JA, Lammers JW, Jove R, Koenderman L . 1999 Blood 94: 1108–1112
Gouilleux F, Wakao H, Mundt M, Groner B . 1994 EMBO J. 13: 4361–4369
Hanke JH, Gardner JP, Dow RL, Changelian PS, Brissette WH, Weringer EJ, Pollok BA, Connelly PA . 1996 J. Biol. Chem. 271: 695–701
Ihle JN . 1995 Adv. Immunol. 60: 1–35
Iwatsuki K, Endo T, Misawa H, Yokouchi M, Matsumoto A, Ohtsubo M, Mori KJ, Yoshimura A . 1997 J. Biol. Chem. 272: 8149–8152
Kazansky AV, Kabotyanski EB, Wyszomierski SL, Mancini MA, Rosen JM . 1999 J. Biol. Chem. 274: 22484–22492
Kitanaka A, Waki M, Kamano H, Tanaka T, Kubota Y, Ohnishi H, Takahara J, Irino S . 1994 Biochem. Biophys. Res. Commun. 201: 1534–1540
Kitanaka A, Mano H, Conley ME, Campana D . 1998 Blood 91: 940–948
Kitanaka A, Suzuki T, Ito C, Nishigaki H, Coustan-Smith E, Tanaka T, Kubota Y, Campana D . 1999 J. Immunol. 162: 1952–1958
Liu X, Robinson GW, Gouilleux F, Groner B, Hennighausen L . 1995 Proc. Natl. Acad. Sci. USA 92: 8831–8835
Meydan N, Grunberger T, Dadi H, Shahar M, Arpaia E, Lapidot Z, Leeder JS, Freedman M, Cohen A, Gazit A, Levitzki A, Roifman CM . 1996 Nature 379: 645–648
Mui AL, Wakao H, O'Farrell AM, Harada N, Miyajima A . 1995 EMBO J. 14: 1166–1175
Mukhopadhyay D, Tsiokas L, Zhou XM, Foster D, Brugge JS, Sukhatme VP . 1995 Nature 375: 577–581
Nosaka T, Kawashima T, Misawa K, Ikuta K, Mui AL, Kitamura T . 1999 EMBO J. 18: 4754–4765
Olayioye MA, Beuvink I, Horsch K, Daly JM, Hynes NE . 1999 J. Biol. Chem. 274: 17209–17218
O'Shea JJ . 1997 Immunity 7: 1–11
Reddy EP, Korapati A, Chaturvedi P, Rane S . 2000 Oncogene 19: 2532–2547
Socolovsky M, Fallon AE, Wang S, Brugnara C, Lodish HF . 1999 Cell 98: 181–191
Teglund S, McKay C, Schuetz E, van Deursen JM, Stravopodis D, Wang D, Brown M, Bodner S, Grosveld G, Ihle JN . 1998 Cell 93: 841–850
Thomas SM, Brugge JS . 1997 Annu. Rev. Cell Dev. Biol. 13: 513–609
Tilbrook PA, Ingley E, Williams JH, Hibbs ML, Klinken SP . 1997 EMBO J. 16: 1610–1619
van der Geer P, Hunter T, Lindberg RA . 1994 Annu. Rev. Cell Dev. Biol. 10: 251–337
Wakao H, Harada N, Kitamura T, Mui AL, Miyajima A . 1995 EMBO J. 14: 2527–2535
Yu CL, Meyer DJ, Campbell GS, Larner AC, Carter-Su C, Schwartz J, Jove R . 1995 Science 269: 81–83
Acknowledgements
We would like to thank Dr T Kitamura for providing the STAT5 cDNAs, Dr JN Ihle for providing the JAK2 cDNA and Dr M Kotb for a critical reading of the manuscript. We would also like to thank Kirin Brewery for providing recombinant human EPO.
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Okutani, Y., Kitanaka, A., Tanaka, T. et al. Src directly tyrosine-phosphorylates STAT5 on its activation site and is involved in erythropoietin-induced signaling pathway. Oncogene 20, 6643–6650 (2001). https://doi.org/10.1038/sj.onc.1204807
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DOI: https://doi.org/10.1038/sj.onc.1204807
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