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Altered affinity of CBFβ-SMMHC for Runx1 explains its role in leukemogenesis

Nature Structural Biology volume 9, pages 674–679 (2002)Cite this article

Abstract

Chromosomal translocations involving the human CBFB gene, which codes for the non-DNA binding subunit of CBF (CBFβ), are associated with a large percentage of human leukemias. The translocation inv(16) that disrupts the CBFB gene produces a chimeric protein composed of the heterodimerization domain of CBFβ fused to the C-terminal coiled-coil domain from smooth muscle myosin heavy chain (CBFβ-SMMHC). Isothermal titration calorimetry results show that this fusion protein binds the Runt domain from Runx1 (CBFα) with higher affinity than the native CBFβ protein. NMR studies identify interactions in the CBFβ portion of the molecule, as well as the SMMHC coiled-coil domain. This higher affinity provides an explanation for the dominant negative phenotype associated with a knock-in of the CBFB-MYH11 gene and also helps to provide a rationale for the leukemia-associated dysregulation of hematopoietic development that this protein causes.

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Figure 1: ITC measurements of the binding of CBFβ and CBFβ-SMMHC proteins to the Runx1 Runt domain (amino acids 41–190).
Figure 2: Saturation enthalpies per mole of Runt domain for CBFβ (red), CBFβ-SMMHC47 (blue), 2:1 mixture of CBFβ/CBFβ-SMMHC47 (green) and a 4:1 mixture of CBFβ/CBFβ-SMMHC47 (orange) to the Runx1 Runt domain, determined by ITC measurements.
Figure 3: Results of saturation transfer experiment performed on a 1:1 complex of triply labeled CBFβ-SMMHC47/unlabeled Runt domain.
Figure 4: ITC measurements of binding for mutated proteins and in the presence of DNA.
Figure 5

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Acknowledgements

This work was supported by grants from the United States Public Health Service (NIH) to J.H.B. The National High Magnetic Field Laboratory at Florida State University is supported by NSF and the State of Florida. NMRFAM is supported by the NIH Biomedical Technology Program and NIH Shared Instrumentation Program. R.L.B. and N.A.S. are supported by grants from the United States Public Health Service (NIH). M.K. is supported by a fellowship from the Damon Runyon Cancer Research Foundation.

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Authors and Affiliations

  1. Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, 22908, Virginia, USA

    Stephen M. Lukasik, Takeshi Corpora, Sarah Tomanicek, Yuanhong Li & John H. Bushweller

  2. Department of Biochemistry, Dartmouth Medical School, Hanover, 03755, New Hampshire, USA

    Lina Zhang & Nancy A. Speck

  3. National Human Genome Research Institute, National Institutes of Health, Bethseda, 20892, Maryland, USA

    Mondira Kundu & P. Paul Liu

  4. Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham, 03824, New Hampshire, USA

    Kari Hartman & Thomas M. Laue

  5. Department of Pharmacology, University of Virginia, Charlottesville, 22908, Virginia, USA

    Rodney L. Biltonen

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Lukasik, S., Zhang, L., Corpora, T. et al. Altered affinity of CBFβ-SMMHC for Runx1 explains its role in leukemogenesis. Nat Struct Mol Biol 9, 674–679 (2002). https://doi.org/10.1038/nsb831

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