The structural basis of protein acetylation by the p300/CBP transcriptional coactivator

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Abstract

The transcriptional coactivator p300/CBP (CREBBP) is a histone acetyltransferase (HAT) that regulates gene expression by acetylating histones and other transcription factors. Dysregulation of p300/CBP HAT activity contributes to various diseases including cancer1,2,3,4. Sequence alignments, enzymology experiments and inhibitor studies on p300/CBP have led to contradictory results about its catalytic mechanism and its structural relation to the Gcn5/PCAF and MYST HATs5,6,7,8,9. Here we describe a high-resolution X-ray crystal structure of a semi-synthetic heterodimeric p300 HAT domain in complex with a bi-substrate inhibitor, Lys-CoA. This structure shows that p300/CBP is a distant cousin of other structurally characterized HATs, but reveals several novel features that explain the broad substrate specificity and preference for nearby basic residues. Based on this structure and accompanying biochemical data, we propose that p300/CBP uses an unusual ‘hit-and-run’ (Theorell–Chance) catalytic mechanism that is distinct from other characterized HATs. Several disease-associated mutations can also be readily accounted for by the p300 HAT structure. These studies pave the way for new epigenetic therapies involving modulation of p300/CBP HAT activity.

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Figure 1: Structure of p300 HAT domain and comparison of the p300 HAT domain with other HATs.
Figure 2: Lys-CoA recognition by p300.
Figure 3: Substrate-binding surface and mutagenesis studies.
Figure 4: Catalytic mechanism of p300 HAT.

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Protein Data Bank

Data deposits

The structure of the p300 HAT–Lys-CoA complex has been submitted to the Protein Data Bank under accession number 3BIY.

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Acknowledgements

We appreciate advice on the manuscript from W. Cleland, J. Stivers, A. Mildvan and D. Leahy. We thank the National Institutes of Health and the FAMRI, Kaufman and Keck foundations for financial support.

Author Contributions X.L. and L.W. designed and performed reported experiments and prepared manuscript figures and text; K.Z. and P.R.T. developed key reagents and performed preliminary studies that led to the reported experiments; Y.H. developed a key reagent and designed and interpreted some of the experiments. R.M. and P.A.C. designed and supervised experiments and prepared manuscript text. All authors read and approved the submitted manuscript.

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Correspondence to Ronen Marmorstein or Philip A. Cole.

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Supplementary Information

The file contains Supplementary Methods with additional references; Supplementary Figures S1-S10 with Legends and Supplementary Tables S1-S3. (PDF 1922 kb)

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Liu, X., Wang, L., Zhao, K. et al. The structural basis of protein acetylation by the p300/CBP transcriptional coactivator. Nature 451, 846–850 (2008) doi:10.1038/nature06546

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