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Core-binding factor β interacts with Runx2 and is required for skeletal development

Nature Genetics volume 32, pages 633–638 (2002)Cite this article

Abstract

Core-binding factor β (CBFβ, also called polyomavirus enhancer binding protein 2β (PEBP2B)) is associated with an inversion of chromosome 16 and is associated with acute myeloid leukemia in humans1. CBFβ forms a heterodimer with RUNX1 (runt-related transcription factor 1), which has a DNA binding domain homologous to the pair-rule protein runt in Drosophila melanogaster. Both RUNX1 and CBFβ are essential for hematopoiesis2,3,4,5,6. Haploinsufficiency of another runt-related protein, RUNX2 (also called CBFA1), causes cleidocranial dysplasia in humans7 and is essential in skeletal development by regulating osteoblast differentiation and chondrocyte maturation8,9,10,11,12,13,14,15. Mice deficient in Cbfb (Cbfb−/−) die at midgestation4,5,6, so the function of Cbfβ in skeletal development has yet to be ascertained. To investigate this issue, we rescued hematopoiesis of Cbfb−/− mice by introducing Cbfb using the Gata1 promoter. The rescued Cbfb−/− mice recapitulated fetal liver hematopoiesis in erythroid and megakaryocytic lineages and survived until birth, but showed severely delayed bone formation. Although mesenchymal cells differentiated into immature osteoblasts, intramembranous bones were poorly formed. The maturation of chondrocytes into hypertrophic cells was markedly delayed, and no endochondral bones were formed. Electrophoretic mobility shift assays and reporter assays showed that Cbfβ was necessary for the efficient DNA binding of Runx2 and for Runx2-dependent transcriptional activation. These findings indicate that Cbfβ is required for the function of Runx2 in skeletal development.

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Figure 1: Generation of Cbfb−/−tg mice.
Figure 2: Hematopoiesis in Cbfb−/−tg mice at E18.5.
Figure 3: Examination of the skeletal system in wildtype and Cbfb−/−tg embryos at E18.5.
Figure 4: Intramembranous ossification in craniofacial bones.
Figure 5: Endochondral ossification in limbs.
Figure 6: Requirement of Cbfβ for osteoblast differentiation, DNA binding of Runx2 and Runx2-dependent transcriptional activation.

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Acknowledgements

We thank M. Yamamoto for the Gata1 promoter; Y. Ito for antibodies against Runx2 and Cbfβ; T. Kitamura for retroviral vector and Platinum-E; H. Harada for Runx2 and Cbfb cDNA; Y. Fujio for pACCMV.pLpA vector; A. Yamaguchi and M. Iwamoto for critically reading this manuscript; K. Sasaki, S. Bae and H. Enomoto for technical advice; R. Hiraiwa for maintaining mouse colonies; and M. Yanagita for secretarial assistance. This work was supported by grants from the Japanese Ministry of Education, Culture, Sports, Science and Technology.

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Authors and Affiliations

  1. Department of Molecular Medicine, Osaka University Medical School, 2-2 Yamada-oka, Suita, 565-0871, Osaka, Japan

    Carolina A. Yoshida, Tatsuya Furuichi, Ryo Fukuyama, Naoko Kanatani, Shinji Kobayashi & Toshihisa Komori

  2. Department of Orthodontics and Dentofacial Orthopedics, Osaka University Faculty of Dentistry, Suita, Osaka, Japan

    Carolina A. Yoshida & Kenji Takada

  3. Department of Pharmacology, Faculty of Pharmaceutical Science, Setsunan University, Hirakata, Osaka, Japan

    Takashi Fujita & Ryo Fukuyama

  4. Teijin Institute for Biomedical Research, Teijin, Hino, Tokyo, Japan

    Shinji Kobayashi

  5. Department of Molecular Immunology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan

    Masanobu Satake

  6. Japan Science and Technology Corporation, Japan

    Toshihisa Komori

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  1. Carolina A. Yoshida
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Correspondence to Toshihisa Komori.

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Yoshida, C., Furuichi, T., Fujita, T. et al. Core-binding factor β interacts with Runx2 and is required for skeletal development. Nat Genet 32, 633–638 (2002). https://doi.org/10.1038/ng1015

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