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Proteomic identification of C/EBP-DBD multiprotein complex: JNK1 activates stem cell regulator C/EBPα by inhibiting its ubiquitination

Oncogene volume 26, pages 1789–1801 (2007)Cite this article

Abstract

Functional inactivation of transcription factors in hematopoietic stem cell development is involved in the pathogenesis of acute myeloid leukemia (AML). Stem cell regulator C/enhancer binding protein (EBP)α is among such transcription factors known to be inactive in AML. This is either due to mutations or inhibition by protein–protein interactions. Here, we applied a mass spectrometry-based proteomic approach to systematically identify putative co-activator proteins interacting with the DNA-binding domain (DBD) of C/EBP transcription factors. In our proteomic screen, we identified c-Jun N-terminal kinase (JNK) 1 among others such as PAK6, MADP-1, calmodulin-like skin proteins and ZNF45 as proteins interacting with DBD of C/EBPs from nuclear extract of myelomonocytic U937 cells. We show that kinase JNK1 physically interacts with DBD of C/EBPα in vitro and in vivo. Furthermore, we show that active JNK1 inhibits ubiquitination of C/EBPα possibly by phosphorylating in its DBD. Consequently, JNK1 prolongs C/EBPα protein half-life leading to its enhanced transactivation and DNA-binding capacity. In certain AML patients, however, the JNK1 mRNA expression and its kinase activity is decreased which suggests a possible reason for C/EBPα inactivation in AML. Thus, we report the first proteomic screen of C/EBP-interacting proteins, which identifies JNK1 as positive regulator of C/EBPα.

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Acknowledgements

We thank Dr Roger Davis for providing pCDNA3-MKK7, pCDNA3-MEKK1-flag; Dr Daniel G Tenen and Dr Claus Nerlov for GST, GST-DBD and GST-C/EBPα expressin plasmids. This work was supported by Deutsche José Carreras Leukamie-Stiftung e.V. grant, DJCLS F03/03 to Dr Trivedi AK.

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Authors and Affiliations

  1. Department of Internal Medicine IV – Hematology and Oncology, Bone Marrow Transplantation Section, State Center for Cell and Gene Therapy, Martin-Luther-University Halle-Wittenberg, Halle, SA, Germany

    A K Trivedi, D Bararia, M Christopeit, A A PeerZada, S M Singh & G Behre

  2. Department of Gene Vectors, GSF-National Research Center for Environment and Health, Munich, Germany

    A Kieser

  3. Department of Internal Medicine III Grosshadern, University of Munich, Munich, Germany

    W Hiddemann

  4. Andrology Section, Martin-Luther-University Halle-Wittenberg, Halle, Germany

    H M Behre

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  1. A K Trivedi
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  2. D Bararia
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Correspondence to G Behre.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Trivedi, A., Bararia, D., Christopeit, M. et al. Proteomic identification of C/EBP-DBD multiprotein complex: JNK1 activates stem cell regulator C/EBPα by inhibiting its ubiquitination. Oncogene 26, 1789–1801 (2007). https://doi.org/10.1038/sj.onc.1209964

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