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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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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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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DOI: https://doi.org/10.1038/nsb936
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