Specific Acetylation Patterns of H2A.Z Form Transient Interactions with the BPTF Bromodomain

Gabriella T. Perell, Neeraj K. Mishra, Babu Sudhamalla, Peter D. Ycas, Kabirul Islam, William C.K. Pomerantz

Research output: Contribution to journalArticle

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Abstract

Post-translational lysine acetylation of histone tails affects both chromatin accessibility and recruitment of multifunctional bromodomain-containing proteins for modulating transcription. The bromodomain- and PHD finger-containing transcription factor (BPTF) regulates transcription but has also been implicated in high gene expression levels in a variety of cancers. In this report, the histone variant H2A.Z, which replaces H2A in chromatin, is evaluated for its affinity for BPTF with a specific recognition pattern of acetylated lysine residues of the N-terminal tail region. Although BPTF immunoprecipitates H2A.Z-containing nucleosomes, a direct interaction with its bromodomain has not been reported. Using protein-observed fluorine nuclear magnetic resonance (PrOF NMR) spectroscopy, we identified a diacetylation of H2A.Z on lysine residues 4 and 11, with the highest affinity for BPTF with a Kd of 780 μM. A combination of subsequent 1H NMR Carr-Purcell-Meiboom-Gill experiments and photo-cross-linking further confirmed the specificity of the diacetylation pattern at lysines 4 and 11. Because of an adjacent PHD domain, this transient interaction may contribute to a higher-affinity bivalent interaction. Further evaluation of specificity toward a set of bromodomains, including two BET bromodomains (Brd4 and BrdT) and two Plasmodium falciparum bromodomains, resulted in one midmicromolar affinity binder, PfGCN5 (Kd = 650 μM). With these biochemical experiments, we have identified a direct interaction of histone H2A.Z with bromodomains with a specific acetylation pattern that further supports the role of H2A.Z in epigenetic regulation.

Original languageEnglish (US)
Pages (from-to)4607-4615
Number of pages9
JournalBiochemistry
Volume56
Issue number35
DOIs
StatePublished - Sep 5 2017

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Acetylation
Lysine
Transcription Factors
Histones
Transcription
Chromatin
Fluorine
Nucleosomes
Plasmodium falciparum
Epigenomics
Gene expression
Nuclear magnetic resonance spectroscopy
Pattern recognition
Binders
Proteins
Magnetic Resonance Spectroscopy
Experiments
Nuclear magnetic resonance
Gene Expression
fetal Alzheimer antigen

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Specific Acetylation Patterns of H2A.Z Form Transient Interactions with the BPTF Bromodomain. / Perell, Gabriella T.; Mishra, Neeraj K.; Sudhamalla, Babu; Ycas, Peter D.; Islam, Kabirul; Pomerantz, William C.K.

In: Biochemistry, Vol. 56, No. 35, 05.09.2017, p. 4607-4615.

Research output: Contribution to journalArticle

Perell, Gabriella T. ; Mishra, Neeraj K. ; Sudhamalla, Babu ; Ycas, Peter D. ; Islam, Kabirul ; Pomerantz, William C.K. / Specific Acetylation Patterns of H2A.Z Form Transient Interactions with the BPTF Bromodomain. In: Biochemistry. 2017 ; Vol. 56, No. 35. pp. 4607-4615.
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