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Structural basis for negative regulation of hypoxia-inducible factor-1α by CITED2

Nature Structural & Molecular Biology volume 10, pages 504–512 (2003)Cite this article

Abstract

Expression of hypoxia-responsive genes is mediated by the heterodimeric transcription factor hypoxia-inducible factor-1 (HIF-1) in complex with the p300/CREB-binding protein (p300/CBP) transcriptional coactivator. The protein CITED2, which binds p300/CBP, is thought to be a negative regulator of HIF-1 transactivation. We show that the CITED2 transactivation domain (TAD) disrupts a complex of the HIF-1α C-terminal TAD (C-TAD) and the cysteine-histidine–rich 1 (CH1) domain of p300/CBP by binding CH1 with high affinity. The high-resolution solution structure of the CITED2 TAD–p300 CH1 complex shows that the CITED2 TAD, like the HIF-1α C-TAD, folds on a helical, Zn2+-containing CH1 scaffold. The CITED2 TAD binds a different, more extensive surface of CH1 than does the HIF-1α C-TAD. However, a conserved 'LPXL' sequence motif in CITED2 and HIF-1α interacts with an overlapping binding site on CH1. Mutation of the LPEL sequence in full-length CITED2 abolishes p300 binding in vivo. These findings reveal that CITED2 regulates HIF-1 by competing for a hot spot on the p300 CH1 domain.

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Figure 1: Homologous sequences of CITED2 TAD and p300 CH1 domain.
Figure 2: Binding of CITED2 to p300 and stability of the CITED2–p300 complex.
Figure 3: Structure of the CITED2–p300 complex and comparison to the HIF-1α–p300 complex.
Figure 4: Contact maps of the CITED2–p300 complex compared to the HIF-1α–p300 complex.
Figure 5: Intermolecular contacts between CITED2 and p300.
Figure 6: Transcription factor binding surfaces on CH1.
Figure 7: Binding of an LPEL mutant of CITED2 in vivo.

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Acknowledgements

This work was supported in part by grants from the Dana-Farber Cancer Institute–Novartis Drug Discovery Program to M.J.E. and A.L.K., and a US National Science Foundation grant to G.W. M.J.E. is a recipient of a scholar award from the Leukemia and Lymphoma Society. S.J.F. is a recipient of a National Institutes of Health K08 award from the National Heart, Lung, and Blood Institute, and an ASH Scholar Award from the American Society of Hematology.

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Authors and Affiliations

  1. Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, 41 Avenue Louis Pasteur, Boston, 02115, Massachusetts, USA

    Steven J Freedman

  2. Department of Biological Chemistry and Molecular Pharmacology, 240 Longwood Avenue, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Zhen-Yu J Sun, Gerhard Wagner & Michael J Eck

  3. Department of Cancer Biology, Dana-Farber Cancer Institute, 44 Binney Street, Boston, 02215, Massachusetts, USA

    Andrew L Kung & Michael J Eck

  4. Oncology Department, Novartis Pharmaceuticals Corporation, One Health Plaza, East Hanover, 07936, New Jersey, USA

    Dennis S France

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Correspondence to Michael J Eck.

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Freedman, S., Sun, ZY., Kung, A. et al. Structural basis for negative regulation of hypoxia-inducible factor-1α by CITED2. Nat Struct Mol Biol 10, 504–512 (2003). https://doi.org/10.1038/nsb936

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