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Oncogenes Fusion Genes and Tumor Suppressor Genes

Proteomic discovery of Max as a novel interacting partner of C/EBPα: a Myc/Max/Mad link

Leukemia volume 20, pages 2137–2146 (2006)Cite this article

Abstract

The transcription factor CCAAT/enhancer binding protein a (C/EBPα) is important in the regulation of granulopoiesis and is disrupted in human acute myeloid leukemia. In the present study, we sought to identify novel C/EBPα interacting proteins in vivo through immunoprecipitation using mass spectrometry-based proteomic techniques. We identified Max, a heterodimeric partner of Myc, as one of the interacting proteins of C/EBPα in our screen. We confirmed the in vivo interaction of C/EBPα with Max and showed that this interaction involves the basic region of C/EBPα. Endogenous C/EBPα and Max, but not Myc and Max, colocalize in intranuclear structures during granulocytic differentiation of myeloid U937 cells. Max enhanced the transactivation capacity of C/EBPα on a minimal promoter. A chromatin immunoprecipitation assay revealed occupancy of the human C/EBPα promoter in vivo by Max and Myc under cellular settings and by C/EBPα and Max under retinoic acid induced granulocytic differentiation. Interestingly, enforced expression of Max and C/EBPα results in granulocytic differentiation of the human hematopoietic CD34+ cells, as evidenced by CD11b, CD15 and granulocyte colony-stimulating factor receptor expression. Silencing of Max by short hairpin RNA in CD34+ and U937 cells strongly reduced the differentiation-inducing potential of C/EBPα, indicating the importance of C/EBPα–Max in myeloid progenitor differentiation. Taken together, our data reveal Max as a novel co-activator of C/EBPα functions, thereby suggesting a possible link between C/EBPα and Myc–Max–Mad network.

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Acknowledgements

We thank Dr Dirk Eick for providing Max expression plasmids, Dr Robert Eisenman for in vitro-translatable Max plasmid and Dr Alan Friedman for C/EBPα constructs. This work was financially supported by Grant F03/04 of the Deutsche-Jose-Carreras Leukemia Stiftung, Muenchen Germany to Peer Zada AA.

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Authors and Affiliations

  1. Bone Marrow Transplantation Unit, State Center for Cell and Gene Therapy, Clinic Internal Medicine IV, Martin-Luther-University, Halle, Germany

    A A Zada, J A Pulikkan, D Bararia, M Geletu, A K Trivedi, M Y Balkhi & G Behre

  2. Department of Internal Medicine III, University Hospital Grosshadern, Munich, Germany

    W D Hiddemann

  3. Division of Hematology/Oncology, Harvard Institutes of Medicine, Boston, MA, USA

    D G Tenen

  4. Andrology Section, Clinic Krollwitz, Martin-Luther-University, Halle, Germany

    H M Behre

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  1. A A Zada
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Correspondence to G Behre.

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

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Zada, A., Pulikkan, J., Bararia, D. et al. Proteomic discovery of Max as a novel interacting partner of C/EBPα: a Myc/Max/Mad link. Leukemia 20, 2137–2146 (2006). https://doi.org/10.1038/sj.leu.2404438

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