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A functional SUMO-interacting motif in the transactivation domain of c-Myb regulates its myeloid transforming ability

Oncogene volume 30, pages 212–222 (2011)Cite this article

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Abstract

c-Myb is an essential hematopoietic transcription factor that controls proliferation and differentiation of progenitors during blood cell development. Whereas sumoylation of the C-terminal regulatory domain (CRD) is known to have a major impact on the activity of c-Myb, no role for noncovalent binding of small ubiquitin-like modifier (SUMO) to c-Myb has been described. Based on the consensus SUMO-interacting motif (SIM), we identified and examined putative SIMs in human c-Myb. Interaction and reporter assays showed that the SIM in the in the transactivation domain of c-Myb (V267NIV) is functional. This motif is necessary for c-Myb to be able to interact noncovalently with SUMO, preferentially SUMO2/3. Destroying the SUMO-binding properties by mutation resulted in a large increase in the transactivation potential of c-Myb. Mutational analysis and overexpression of conjugation-defective SUMO argued against intramolecular repression caused by sumoylated CRD and in favor of SUMO-dependent repression in trans. Using both a myeloid cell line-based assay and a primary hematopoietic cell assay, we addressed the transforming abilities of SUMO binding and conjugation mutants. Interestingly, only loss of SUMO binding, and not SUMO conjugation, enhanced the myeloid transformational potential of c-Myb. c-Myb with the SIM mutated conferred a higher proliferative ability than the wild-type and caused an effective differentiation block. This establishes SUMO binding as a mechanism involved in modulating the transactivation activity of c-Myb, and responsible for keeping the transforming potential of the oncoprotein in check.

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Acknowledgements

We thank Marit Ledsaak for excellent technical assistance and Professor D Livingston, Professor G Del Sal, Professor RT Hay, Professor R Grosschedl and Professor ET Yeh for providing us with relevant expression constructs. This work was supported by the Norwegian Cancer Society (TS) and the FUGE program of the Norwegian Research Council. DRP was supported by a scholarship from The Leukaemia Foundation (Australia).

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  1. T Sæther and D R Pattabiraman: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biosciences, University of Oslo, Oslo, Norway

    T Sæther, A H Alm-Kristiansen, L T Vogt-Kielland & O S Gabrielsen

  2. University of Queensland Diamantina Institute, Brisbane, Queensland, Australia

    D R Pattabiraman & T J Gonda

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  1. T Sæther
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Correspondence to T Sæther.

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Sæther, T., Pattabiraman, D., Alm-Kristiansen, A. et al. A functional SUMO-interacting motif in the transactivation domain of c-Myb regulates its myeloid transforming ability. Oncogene 30, 212–222 (2011). https://doi.org/10.1038/onc.2010.397

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