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Identification and Characterization of Constitutively Active Stat5

  • Tetsuya Nosaka
  • Mayumi Onishi
  • Toshiyuki Kawashima
  • Koji Yamada
  • Kazuhide Misawa
  • Kouichi Ariyoshi
  • Masayuki Towatari
  • Hidehiko Saito
  • Kenzaburo Tani
  • Shigetaka Asano
  • Atsushi Miyajima
  • Toshio Kitamura
Chapter

Summary

We have recently identified several constitutively active STAT5s using polymerase chain reaction (PCR)-driven random mutagenesis followed by retrovirus- mediated expression screening. One such mutant STAT5A1*6 harbors two point mutations; one in the effector domain and the other in the DNA binding domain, and induces factor-independent growth of several IL-3-dependent cell lines. The mutant STAT5A1*6 shows constitutive phosphorylation on its tyrosine residues, translocates into the nucleus, binds the target DNA sequence, and stimulates transcription in the absence of IL-3 stimulation. Further biochemical analyses suggested that a molecular basis for the constitutive activity of the mutant STAT5A is the increased stability of the phosphorylated form of STAT5. In addition to the ability of inducing factor- independent growth of IL-3-dependent cell lines, the mutant STAT5A induces apop- tosis or differentiation under some conditions or in a different cell line, implicating STAT5 as a molecular switch for pleiotropic functions of cytokine receptors. The relationship between active STAT5 and leukemogenesis will also be discussed.

Keywords

Random Mutagenesis Hematopoietic Factor Active STAT5s Nonreceptor Protein Tyrosine Kinase Mutant STAT5B 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Taniguchi, T: Cytokine signaling through nonreceptor protein tyrosine kinases. Science 268: 251, 1995.PubMedCrossRefGoogle Scholar
  2. 2.
    Mui, AL-F, Wakao, H, O’Farrell, A, Harada, N, Miyajima, A: Interleukin-3, granulocyte-macrophage colony stimulating factor and interleukin-5 transduce signals through two STAT5 homologs. EMBO J 14: 1166, 1995.PubMedGoogle Scholar
  3. 3.
    Schindler, C, Darnell, J, Jr.: Transcriptional responses to polypeptide ligands: the JAK-STAT pathway. Annu Rev Biochem 64: 621, 1995.PubMedCrossRefGoogle Scholar
  4. 4.
    Ihle, JN: STATs: signal transducers and activators of transcription. Cell 84: 331, 1996.PubMedCrossRefGoogle Scholar
  5. 5.
    O’Shea, JJ: Jaks, STATs, cytokine signal transduction, and immunoregulation: Are we there yet? Immunity 7: 1, 1997.PubMedCrossRefGoogle Scholar
  6. 6.
    Jacobson, NG, Szabo, SJ, Weber-Nordt, RM, Zhong, Z, Schreiber, RD, Darnell, J, Jr., Murphy, KM: Interleukin 12 signaling in T helper type 1 (Th1) cells involves tyrosine phosphorylation of signal transducer and activator of transcription (Stat)3 and Stat4. J Exp Med 181: 1755, 1995.PubMedCrossRefGoogle Scholar
  7. 7.
    Bacon, CM, Petricoin, EF, Ortaldo, JR, Rees, RC, Larner, AC, Johnston, JA, O’Shea, JJ: Interleukin 12 (IL-12) induces tyrosine phosphorylation and activation of STAT4 in human lymphocytes. Proc. Natl. Acad. Sci. USA. 92: 7307, 1995.PubMedCrossRefGoogle Scholar
  8. 8.
    Takeda, K, Kishimoto, T, Akira, S: STAT6: its role in interleukin 4-mediated biological functions. J. Mol. Med. 75: 317, 1997.PubMedCrossRefGoogle Scholar
  9. 9.
    Sato, N, Sakamaki, K, Terada, N, Arai, K, Miyajima, A: Signal transduction by the high-affinity GM-CSF receptor: two distinct cytoplasmic regions of the common beta subunit responsible for different signaling. EMBO J 12: 4181, 1993.PubMedGoogle Scholar
  10. 10.
    Kinoshita, T, Yokota, T, Arai, K, Miyajima, A: Suppression of apoptotic death in hematopoietic cells by signalling through the IL-3/GM-CSF receptors. EMBO J 14: 266, 1995.PubMedGoogle Scholar
  11. 11.
    Mui, ALF, Wakao, H, Kinoshita, T, Kitamura, T, Miyajima, A: Suppression of interleukin-3-induced gene expression by a C-terminal truncated Stat5: role of Stat5 in proliferation. EMBO J 15: 2425, 1996.PubMedGoogle Scholar
  12. 12.
    Damen, JE, Wakao, H, Miyajima, A, Krosl, J, Humphries, RK, Cutler, RL, Krystal, G: Tyrosine 343 in the erythropoietin receptor positively regulates erythropoietin-induced cell proliferation and Stat5 activation. EMBO J 14: 5557, 1995.PubMedGoogle Scholar
  13. 13.
    Friedmann, MC, Migone, T, Russell, SM, Leonard, WJ: Different interleukin 2 receptor β-chain tyrosines couple to at least two signaling pathways and synergistically mediate interleukin 2-induced proliferation. Proc. Natl. Acad. Sci. USA 93: 2077, 1996.PubMedCrossRefGoogle Scholar
  14. 14.
    Gobert, S, Chretien, S, Gouilleux, F, Muller, O, Pallard, C, Dusanter-Fourt, I, Groner, B, Lacombe, C, Gisselbrecht, S, Mayeux, P: Identification of tyrosine residues within the intracellular domain of the erythropoietin receptor crucial for STAT5 activation. EMBO J. 15: 2434, 1996.PubMedGoogle Scholar
  15. 15.
    Iwatsuki, K, Endo, T, Misawa, H, Yokouchi, M, Matsumoto, A, Ohtsubo, M, Mori, KJ, Yoshimura, A: STAT5 activation correlates with erythropoietin-mediated erythroid differentiation of an erythroleukemia cell line. J. Biol. Chem. 272: 8149, 1997.PubMedCrossRefGoogle Scholar
  16. 16.
    Wakao, H, Chida, D, Damen, JE, Krystal, G, Miyajima, A: A possible involvement of Stat5 in erythropoietin-induced hemoglobin synthesis. Biochem. Byophys. Res. Commun. 234: 198, 1997.CrossRefGoogle Scholar
  17. 17.
    Fujii, H, Nakagawa, Y, Schindler, U, Kawahara, A, Mori, H, Goulilleux, F, Groner. B, Ihle, JN, Minami, Y, Miyazaki, T, Taniguchi, T: Activation of Stat5 by interleukin 2 requires a carboxyl-terminal region of the interleukin 2 receptor β chain but is not essential for the proliferative signal transmission. Proc. Natl. Acad. Sci. USA. 92: 5482, 1995.PubMedCrossRefGoogle Scholar
  18. 18.
    Quelle, FW, Wang, D, Nosaka, T, Thierfelder, WE, Stravopodis, D, Weinstein, Y Ihle, JN: Erythropoietin induces activation of Stat5 through association with specific tyrosines on the receptor that are not required for a mitogenic response. Mol. Cell. Biol. 16: 1622, 1996.PubMedGoogle Scholar
  19. 19.
    Liu, X, Robinson, GW, Wagner, K-U, Garrett, L, Wynshaw-Boris, A, Hennighausen, L: Stat5a is mandatory for adult mammary gland and lactogenesis. Genes & Dev. 11: 179, 1997.CrossRefGoogle Scholar
  20. 20.
    Udy, GB, Towers, RP, Snell, RG, Wilkins, RJ, Park, S-H, Ram, P, Waxman, DJ, Davey, HW: Requirement of Stat5b for sexual dimorphism of body growth rates and liver gene expression. Proc. Natl. Acad. Sci. U.S.A. 94: 7239, 1997.PubMedCrossRefGoogle Scholar
  21. 21.
    Teglund, S, Mckay, C, Schuetz, E, van Deursen, JM, Stravopodis, D, Wang, D, Brown, M, Bodner, S, Grosveld, G, Ihle, JN: Stat5a and Stat5b proteins have essential and nonessential, or redundant roles in cytokine responses. Cell 93: 841, 1998.PubMedCrossRefGoogle Scholar
  22. 22.
    Onishi, M, Nosaka, T, Misawa, K, Mui, AL-F, Gorman, D, McMahon, M, Miyajima, A, Kitamura, T: Identification and characterization of a constitutive active STAT5 mutant that promotes cell proliferation. Mol. Cell. Biol. 18: 3871, 1998.PubMedGoogle Scholar
  23. 23.
    Kitamura, T, Onishi, M, Kinoshita, S, Shibuya, A, Miyajima, A, Nolan, GP: Efficient screening of retroviral cDNA expression libraries. Proc. Natl. Acad. Sci. U.S.A. 92: 9146, 1995.PubMedCrossRefGoogle Scholar
  24. 24.
    Onishi, M, Kinoshita, S, Morikawa, Y, Shibuya, A, Phillips, J, Lanier, LL, Gorman, DM, Nolan, GP, Miyajima, A, Kitamura, T: Applications of retrovirus-mediated expression cloning. Exp. Hematol. 24: 324, 1996.Google Scholar
  25. 25.
    Migone, T-S, Lin, J-X, Cereseto, A, Mulloy, JC, O’Shea, JJ, Franchini, G, Leonard. WJ: Constitutive activated Jak-STAT pathway in T cells transformed with HTLV-I. Science 269: 79, 1995.PubMedCrossRefGoogle Scholar
  26. 26.
    Cao, X, Tay, A, Guy, GR, Tan, YH: Activation and association of Stat3 with src in v-src-transformed cell lines. Mol. Cell. Biol. 16: 1595, 1996.PubMedGoogle Scholar
  27. 27.
    Danial, NN, Pernis, A, Rothman, PB: Jak-STAT signaling by the v-abl oncogene. Science 269: 1875, 1995.PubMedCrossRefGoogle Scholar
  28. 28.
    Carlesso, N, Frank, DA, Griffin, JD: Tyrosyl phosphorylation and DNA binding activity of signal transducers and activators of transcription (STAT) proteins in hematopoietic cell lines transformed by Bcr/Abl. J. Exp. Med. 183: 811, 1996.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • Tetsuya Nosaka
    • 1
  • Mayumi Onishi
    • 2
  • Toshiyuki Kawashima
    • 1
  • Koji Yamada
    • 1
  • Kazuhide Misawa
    • 1
  • Kouichi Ariyoshi
    • 1
  • Masayuki Towatari
    • 3
  • Hidehiko Saito
    • 3
  • Kenzaburo Tani
    • 4
  • Shigetaka Asano
    • 4
  • Atsushi Miyajima
    • 5
  • Toshio Kitamura
    • 1
    • 6
  1. 1.Department of Hematopoietic Factors Institute of Medical ScienceUniversity of TokyoJapan
  2. 2.Third Department of Internal MedicineUniversity of TokyoJapan
  3. 3.First Department of Internal MedicineUniversity of NagoyaJapan
  4. 4.Department of Hematology-Oncology Institute of Medical ScienceUniversity of TokyoJapan
  5. 5.Institute of Molecular and BiosciencesUniversity of TokyoJapan
  6. 6.Department of Hematopoietic Factors, Institute of Medical ScienceUniversity of TokyoMinato-kuJapan

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