Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Correspondence
  • Published:

A sensitive, non- radioactive and fast method for detection of JNK/SAPK activity in leukemic T cells

Leukemia volume 14, pages 1859–1860 (2000)Cite this article

Stimuli that stress cells, such as chemotherapeutic drugs stimulate a cytoplasmic signaling system consisting of a cascade of protein kinases including the c-Jun kinase (JNK)/stress-activated protein kinase (SAPK).123 Activation of JNK/SAPKs has been linked to the induction of apoptosis45678910111213 and may be a prognostic parameter for the apoptosis sensitivity of leukemia cells.

In conventional detection methods JNK/SAPKs are immunoprecipitated from cell lysates and an in vitro kinase assay is performed by incubating purified JNK/SAPKs together with GST-cJun protein and 32P-γ-ATP. Since JNK/SAPKs phosphorylate and thereby activate transcription factors such as cJun114 the amount of phosphorylated GST-cJun protein reflects activity of JNK/SAPKs which is detected by SDS-PAGE and autoradiography.15

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2

References

  1. Davis RJ . Signal transduction by the c-Jun N-terminal kinase Biochem Soc Sympl 1999 64: 1–12

    CAS  Google Scholar 

  2. Kyriakis JM . Making the connection: coupling of stress-activated ERK/MAPK (extracellular-signal-regulated kinase/mitogen-activated protein kinase) core signalling modules to extracellular stimuli and biological responses Biochem Soc Symp 1999 64: 29–48

    CAS  PubMed  Google Scholar 

  3. Tibbles LA, Woodgett JR . The stress-activated protein kinase pathways Cell Mol Life Sci 1999 55: 1230–1254

    Article  CAS  Google Scholar 

  4. Chen Z, Seimiya H, Naito M, Mashima T, Kizaki A, Dan S, Imaizumi M . ASK1 mediates apoptotic cell death induced by genotoxic stress Oncogene 1999 18: 173–180

    Article  CAS  Google Scholar 

  5. Faris M, Latinis KM, Kempiak SJ, Koretzky GA, Nel A . Stress-induced Fas ligand expression in T cells is mediated through a MEK Kinase 1-regulated response element in the Fas ligand promoter Mol Cell Biol 1998 18: 5414–5424

    Article  CAS  Google Scholar 

  6. Goillot E, Raingeaud J, Ranger A, Tepper RI, Davis RJ, Harlow E, Sanchez I . Mitogen-activated protein kinase-mediated Fas apoptotic signaling pathway Proc Natl Acad Sci USA 1997 94: 3302–3307

    Article  CAS  Google Scholar 

  7. Herr I, Wilhelm D, Meyer E, Jeremias I, Angel P, Debatin K-M . JNK/SAPK activity contributes to TRAIL-induced apoptosis Cell Death Differ 1999 6: 130–135

    Article  CAS  Google Scholar 

  8. Johnson NL, Gardner AM, Diener KM, Lange-Carter CA, Gleavy J, Jarpe MB, Minden A . Signal transduction pathways regulated by mitogen-activated/extracellular response kinase kinase kinase induce cell death J Biol Chem 1996 271: 3229–3237

    Article  CAS  Google Scholar 

  9. Minden A, Lin A, McMahon M, Lange-Carter C, Derijard B, Davis RJ, Johnson GL, Karin M . Differential activation of ERK and JNK mitogen-activated protein kinases by Raf-1 and MEKK Science 1994 266: 1719–1723

    Article  CAS  Google Scholar 

  10. Verheij M, Bose R, Hua Lin X, Yao B, Jarvis WD, Grant S, Birrer MJ, Szabo E, Zon LI, Kyriakis JM, Haimovitz-Friedman A, Fuks Z, Kolesnick RN . Requirement for ceramide initiated JNK/SAPK signalling in stress-induced apoptosis Nature 1996 380: 75–79

    Article  CAS  Google Scholar 

  11. Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME . Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis Science 1995 270: 1326–1331

    Article  CAS  Google Scholar 

  12. Yang X, Khosravi-Far R, Chang HY, Baltimore D . Daxx, a novel Fas-binding protein that activates JNK and apoptosis Cell 1997 89: 1067–1076

    Article  CAS  Google Scholar 

  13. Zanke BW, Boudreau K, Rubie E, Winnett E, Tibbles LA, Zon L, Kyriakis J, Liu F-F, Woodgett JR . The stress-activated protein kinase pathway mediates cell death following injury induced by cis-platinum, UV irradiation or heat Curr Biol 1996 6: 606–613

    Article  CAS  Google Scholar 

  14. Minden A, Karin M . Regulation and function of the JNK subgroup of MAP kinases Biochim Biophys Acta 1997 1333: F85–104

    CAS  PubMed  Google Scholar 

  15. Hibi M, Lin A, Minden A, Karin M . Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the cjun activation domain Genes Dev 1993 7: 2135–2148

    Article  CAS  Google Scholar 

  16. van Dam H, Wilhelm D, Herr I, Steffen A, Herrlich P, Angel P . ATF-2 is preferentially activated by stress-activated protein kinases to mediate c-Jun induction in response to genotoxic agents EMBO J 1995 14: 1798–1811

    Article  CAS  Google Scholar 

  17. Herr I, Wilhelm D, Boehler T, Angel P, Debatin K-M . Activation of CD95 (APO-1/Fas) signaling by ceramide mediates cancer therapy-induced apoptosis EMBO J 1997 16: 6200–6208

    Article  CAS  Google Scholar 

  18. Herr I, Wilhelm D, Boehler T, Angel P, Debatin K-M . JNK/SAPK activity is not sufficient for anticancer therapy-induced apoptosis involving CD95-L, TRAIL and TNF-a Int J Cancer 1999 80: 417–424

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr P Angel for the GST-cJun expression plasmid. This work was supported by grants from the Deutsche Forschungsgemeinschaft.

Author information

Authors and Affiliations

  1. Division of Molecular Oncology, German Cancer Research Center, Heidelberg

    I Herr, D Krilleke & K-M Debatin

  2. University Children's Hospital, Ulm, Germany

    K-M Debatin

Authors
  1. I Herr
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D Krilleke
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K-M Debatin
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Herr, I., Krilleke, D. & Debatin, KM. A sensitive, non- radioactive and fast method for detection of JNK/SAPK activity in leukemic T cells. Leukemia 14, 1859–1860 (2000). https://doi.org/10.1038/sj.leu.2401911

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2401911

This article is cited by

Search

Advanced search

Quick links