Abstract
The Polybromo (PB) protein functions as a key component of the human PBAF chromatin remodeling complex in regulation of gene transcription. PB is made up of modular domains including six bromodomains that are known as acetyl-lysine binding domains. However, histone-binding specificity of the bromodomains of PB has remained elusive. In this study, we report biochemical characterization of all six PB bromodomains' binding to a suite of lysine-acetylated peptides derived from known acetylation sites on human core histones. We demonstrate that bromodomain 2 of PB preferentially recognizes acetylated lysine 14 of histone H3 (H3K14ac), a post-translational mark known for gene transcriptional activation. We further describe the molecular basis of the selective H3K14ac recognition of bromodomain 2 by solving the protein structures in both the free and bound forms using X-ray crystallography and NMR, respectively.
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Acknowledgements
We wish to acknowledge the use of the NMR facility at the New York Structural Biology Center and the staff at the X6A beamline of the National Synchrotron Light Sources at the Brookhaven National Laboratory for facilitating X-ray data collection. We thank D Lencer and W Shreffler (Mount Sinai School of Medicine) for discussion of peptide microarray, S Chakravarty (Mount Sinai School of Medicine) for discussion on structure-based sequence alignment, and A Plotnikov (Mount Sinai School of Medicine), R Page (Brown University) and V Stojanoff (the Brookhaven National Laboratory) for protein structural analysis using X-ray crystallography techniques. This work was supported by a graduate training program fellowship from the National Cancer Institute (ZC-P) and research grants from the National Institutes of Health (M-MZ). ZC-P dedicates this study in the loving memory of Meg Charlop.
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Charlop-Powers, Z., Zeng, L., Zhang, Q. et al. Structural insights into selective histone H3 recognition by the human Polybromo bromodomain 2. Cell Res 20, 529–538 (2010). https://doi.org/10.1038/cr.2010.43
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DOI: https://doi.org/10.1038/cr.2010.43
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