The intrinsic kinase activity of BRD4 spans its BD2-B-BID domains

Weissman, Jocelyn D.; Singh, Amit Kumar; Devaiah, Ballachanda N.; Schuck, Peter; Larue, Ross C.; Singer, Dinah S.

doi:10.1016/j.jbc.2021.101326

Cited by 12 publications

(16 citation statements)

References 26 publications

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“…This is also similar to the reported results on BRD4-T and was observed for BRDT-T here as well. We believe that the intrinsically disordered region connecting the BDs largely contributes to a higher affinity for dsDNA. , This has also been postulated before as the linker is rich in basic residues and has an A-motif (281–300), which is speculated to phosphorylate the Pol II C-terminal domain (CTD) via direct engagement, though nothing is known about its structure …”

Section: Discussionmentioning

confidence: 54%

Alternative Mechanisms for DNA Engagement by BET Bromodomain-Containing Proteins

et al. 2022

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Epigenetic reader domains regulate chromatin structure and modulate gene expression through the recognition of post-translational modifications on histones. Recently, reader domains have also been found to harbor double-stranded (ds) DNA-binding activity, which is as functionally critical as histone association. Here, we explore the dsDNA recognition of the N-terminal bromodomain of the bromodomain and extra-terminal (BET) protein, BRD4. Using protein-observed 19F NMR, 1H-15N HSQC NMR, electrophoretic mobility shift assays (EMSA), and competitive-inhibition assays, we establish the binding surface of dsDNA and find it to be largely overlapping with the acetylated histone (KAc)-binding site. Rather than engaging in electrostatic contacts, we find dsDNA to interact competitively within the KAc-binding pocket. These interactions are distinct from the highly homologous BET bromodomain, BRDT. Nine additional bromodomains have also been characterized for interacting with dsDNA, including tandem BET bromodomains. Together, these studies help establish a binding model for dsDNA interactions with BRD4 bromodomains and elucidate the chromatin-recognition mechanisms of the BRD4 protein for regulating gene expression.

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Section: Discussionmentioning

confidence: 54%

Alternative Mechanisms for DNA Engagement by BET Bromodomain-Containing Proteins

et al. 2022

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“…Interestingly, BET family proteins have been identified as atypical kinases, which might affect gene transcription also by alternative mechanisms [42]. For example, Brd4 may affect gene transcription by phosphorylation of transcription machinery [43]. Brd4 possesses intrinsic kinase activity and may directly phosphorylate RNA polymerase II, TATA-box binding protein associated factor 7 (TAF7), and positive transcription elongation factor b/cyclin-dependent kinase 9 (PTEFb/CDK9) [44,45].…”

Section: Discussionmentioning

confidence: 99%

The Role of Bromodomain and Extraterminal (BET) Proteins in Controlling the Phagocytic Activity of Microglia In Vitro: Relevance to Alzheimer’s Disease

Matuszewska

Cieślik

Wilkaniec

et al. 2022

IJMS

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The correct phagocytic activity of microglia is a prerequisite for maintaining homeostasis in the brain. In the analysis of mechanisms regulating microglial phagocytosis, we focused on the bromodomain and extraterminal domain (BET) proteins: Brd2, Brd3, and Brd4, the acetylation code readers that control gene expression in cooperation with transcription factors. We used pharmacological (JQ1) and genetic (siRNA) inhibition of BET proteins in murine microglial cell line BV2. Inhibition of BET proteins reduced the phagocytic activity of BV2, as determined by using a fluorescent microspheres-based assay and fluorescently labelled amyloid-beta peptides. Gene silencing experiments demonstrated that all brain-existing BET isoforms control phagocytosis in microglia. From a set of 84 phagocytosis-related genes, we have found the attenuation of the expression of 14: Siglec1, Sirpb1a, Cd36, Clec7a, Itgam, Tlr3, Fcgr1, Cd14, Marco, Pld1, Fcgr2b, Anxa1, Tnf, Nod1, upon BET inhibition. Further analysis of the mRNA level of other phagocytosis-related genes which were involved in the pathomechanism of Alzheimer’s disease demonstrated that JQ1 significantly reduced the expression of Cd33, Trem2, and Zyx. Our results indicate the important role of BET proteins in controlling microglial phagocytosis; therefore, targeting BET may be the efficient method of modulating microglial activity.

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“…Likewise, several residues from MLV-EBM, including L399, I401, V392, L403, and W390, are positioned within the hydrophobic core of the ET domain ( Figure 11 D). In addition to the NMR experiments, affinity pull-down experiments further support that the acidic and hydrophobic residues of the ET domain are critical for the binding and potentially linked to its phosphorylation [ 174 , 192 , 202 ]. Based on the solved structure model, Xing et al [ 203 ] reported that 399 LKIRL 403 is a critical amino acid (or hot-spot) sequence that carries out the MLV-EBM/Brd4-ET interactions.…”

Section: Targeting At the Extra-terminal (Et) Domain Of Brd4mentioning

confidence: 99%

BET Bromodomain Inhibitors: Novel Design Strategies and Therapeutic Applications

Xing

Larue

et al. 2023

Molecules

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The mammalian bromodomain and extra-terminal domain (BET) family of proteins consists of four conserved members (Brd2, Brd3, Brd4, and Brdt) that regulate numerous cancer-related and immunity-associated genes. They are epigenetic readers of histone acetylation with broad specificity. BET proteins are linked to cancer progression due to their interaction with numerous cellular proteins including chromatin-modifying factors, transcription factors, and histone modification enzymes. The spectacular growth in the clinical development of small-molecule BET inhibitors underscores the interest and importance of this protein family as an anticancer target. Current approaches targeting BET proteins for cancer therapy rely on acetylation mimics to block the bromodomains from binding chromatin. However, bromodomain-targeted agents are suffering from dose-limiting toxicities because of their effects on other bromodomain-containing proteins. In this review, we provided an updated summary about the evolution of small-molecule BET inhibitors. The design of bivalent BET inhibitors, kinase and BET dual inhibitors, BET protein proteolysis-targeting chimeras (PROTACs), and Brd4-selective inhibitors are discussed. The novel strategy of targeting the unique C-terminal extra-terminal (ET) domain of BET proteins and its therapeutic significance will also be highlighted. Apart from single agent treatment alone, BET inhibitors have also been combined with other chemotherapeutic modalities for cancer treatment demonstrating favorable clinical outcomes. The investigation of specific biomarkers for predicting the efficacy and resistance of BET inhibitors is needed to fully realize their therapeutic potential in the clinical setting.

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The intrinsic kinase activity of BRD4 spans its BD2-B-BID domains

Cited by 12 publications

References 26 publications

Alternative Mechanisms for DNA Engagement by BET Bromodomain-Containing Proteins

Alternative Mechanisms for DNA Engagement by BET Bromodomain-Containing Proteins

The Role of Bromodomain and Extraterminal (BET) Proteins in Controlling the Phagocytic Activity of Microglia In Vitro: Relevance to Alzheimer’s Disease

BET Bromodomain Inhibitors: Novel Design Strategies and Therapeutic Applications

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