DOCSLIB.ORG

Bromodomain-Containing Protein BRPF1 Is a Therapeutic Target for Liver Cancer

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Bromodomain-Containing Protein BRPF1 Is a Therapeutic Target for Liver Cancer ARTICLE https://doi.org/10.1038/s42003-021-02405-6 OPEN Bromodomain-containing protein BRPF1 is a therapeutic target for liver cancer Carol Lai-Hung Cheng1,2, Felice Hoi-Ching Tsang1,2, Lai Wei1,2, Mengnuo Chen1,2, Don Wai-Ching Chin1,2, Jialing Shen1,2, Cheuk-Ting Law1,2, Derek Lee1,2, Carmen Chak-Lui Wong 1,2, Irene Oi-Lin Ng 1,2 & ✉ Chun-Ming Wong 1,2 Epigenetic deregulation plays an essential role in hepatocellular carcinoma (HCC) progres- sion. Bromodomains are epigenetic “readers” of histone acetylation. Recently, bromodomain inhibitors have exhibited promising therapeutic potential for cancer treatment. Using tran- scriptome sequencing, we identified BRPF1 (bromodomain and PHD finger containing 1) as the 1234567890():,; most significantly upregulated gene among the 43 bromodomain-containing genes in human HCC. BRPF1 upregulation was significantly associated with poor patient survival. Gene ablation or pharmacological inactivation of BRPF1 significantly attenuated HCC cell growth in vitro and in vivo. BRPF1 was involved in cell cycle progression, senescence and cancer stemness. Transcriptome sequencing revealed that BRPF1 is a master regulator controlling the expression of multiple key oncogenes, including E2F2 and EZH2. We demonstrated that BRPF1 activated E2F2 and EZH2 expression by facilitating promoter H3K14 acetylation through MOZ/MORF complex. In conclusion, BRPF1 is frequently upregulated in human HCCs. Targeting BRPF1 may be an approach for HCC treatment. 1 State Key Laboratory of Liver Research, The University of Hong Kong, Pok Fu Lam, Hong Kong. 2 Department of Pathology, Li Ka Shing Faculty of Medicine, ✉ The University of Hong Kong, Pok Fu Lam, Hong Kong. email: [email protected] COMMUNICATIONS BIOLOGY | (2021) 4:888 | https://doi.org/10.1038/s42003-021-02405-6 | www.nature.com/commsbio 1 ARTICLE COMMUNICATIONS BIOLOGY | https://doi.org/10.1038/s42003-021-02405-6 epatocellular carcinoma (HCC) is the most common type leukemogenesis associated with MOZ-TIF2 fusion23. Yet, the of liver cancer, accounting for ~80% of primary liver pathological and functional role of BRPF1 in cancer, especially H 1 cancers . HCC is the sixth most common cancer and the liver carcinogenesis, is still largely unknown. fifth deadliest cancer worldwide2,3. HCC is more commonly seen 3 in men and has a higher incidence rate in developing countries . Results HCC development is driven by the accumulation of genetic and Deregulation of bromodomain-containing genes in human epigenetic alterations. The major causes of HCC are hepatitis B fl HCC. Lysine acetylation is an abundant posttranslational mod- (HBV) and hepatitis C (HCV) viral infections, cirrhosis, a atoxin ification in histone and nonhistone proteins. As an unique reader B1 ingestion, excessive alcohol consumption, and nonalcoholic of protein acetylation, bromodomain-containing proteins are fatty liver disease (NAFLD)4. Though the risk factors for HCC are fi important for recognizing and responding to acetylation marks to relatively well de ned, the diagnosis and prognosis of HCC are regulate various cellular processes24,25. We hypothesized that poor. Symptoms usually appear at the late stage of HCC, and 5 deregulation of bromodomain-containing proteins might con- metastasis is commonly observed in HCC patients . tribute to liver carcinogenesis. We therefore compared the Liver transplantation or surgical resection is not applicable for 6 expression levels of 43 human bromodomain-containing genes patients diagnosed at the late stage of HCC . Molecularly targeted (as listed in the ChromoHub database26) in 16 pairs of HBV- therapy is the major method for the treatment of advanced HCC. associated primary HCC samples and their corresponding non- Sorafenib has long been the only FDA-approved molecularly 7 tumor (NT) liver samples by transcriptome sequencing (Fig. 1a, targeted therapy for HCC . Recently, lenvatinib, regorafenib, and b). We found that about half of bromodomain-containing genes cabozantinib have also been approved for HCC treatment. These were highly upregulated in primary HCC. The results showed drugs are all multi-tyrosine kinase inhibitors, which are not that upregulation of bromodomain-containing genes is a com- fi 8–10 speci c and extend the survival rate only by a few months . mon phenomenon in human HCC, implying that the demand for Drug resistance to sorafenib is commonly observed in HCC reading acetylation marks is increased for HCC cell growth. patients11. Immunotherapy maybe a promising new direction for HCC treatment. However, only a subset of HCC patients are 11 Frequent upregulation of BRPF1 in human HCC. In the above responsive to anti-PD1 monoclonal antibodies . Therefore, the fi discovery of potential therapeutic target and the development of transcriptome analysis, we identi ed BRPF1 as the most sig- fi nificantly upregulated bromodomain-containing gene in HCC, new small molecule inhibitors with high speci city for HCC fi treatment are still urgently needed. according to the statistical signi cance (Rank 1) and expression Bromodomain inhibitors represent a novel type of epigenetic change (Rank 10) (Fig. 1b, c). In fact, not just in the in-house drug and hold great promise for cancer therapy. Bromodomains samples, BRPF1 was also upregulated in HCC, colorectal cancer are epigenetic “reader” domains that specifically recognize and kidney cancer from TCGA database (Fig. 1d). The BRPF1 acetylated lysine residues on histones or nonhistone proteins12. upregulation in TCGA HCC cohort was relatively modest com- Lysine acetylation is crucial for gene transcription, nucleosome pared to that in the in-house samples. However, the in-house data assembly, protein–protein interaction and cellular signaling13. comes from Chinese patients associated with HBV-infection, There are 43 bromodomain-containing proteins in humans, while TCGA database contains samples with different aetiological which are divided into eight subgroups depending on their backgrounds and ethnicities. The upregulation of BRPF1 structural similarities. Among these proteins, the members of the expression in HCC tumor samples may be affected by different bromodomain and extra-terminal (BET) family have received aetiological factors. After all, these expression data from TCGA considerable attention14. As registered in ClinicalTrials.gov, there database reinforced our initial observation in the RNA-seq ana- lysis, suggesting that BRPF1 may be essential for cancer devel- are currently 23 ongoing clinical trials of bromodomain inhibitors fi at various phases. These trials largely focus on the therapeutic opment. We found that copy number gain or ampli cation of the effect of BET inhibitors on human cancers, such as acute leuke- BRPF1 gene located on chromosome 3p25 was frequent in human HCC. In addition, HCC with BRPF1 gene copy number gain/ mia and prostate cancer, as well as metabolic disorders. However, fi the implications of other bromodomain-containing proteins in ampli cation had higher BRPF1 mRNA expression than other human carcinogenesis remain to be explored. HCC, suggesting that gain of gene copy number may directly Herein, we identified bromodomain and PHD finger containing contribute to BRPF1 upregulation in human HCC (Fig. 1e). 1(BRPF1) as the most significantly upregulated bromodomain- BRPF1, an epigenetic reader with multiple reader domains containing gene in human HCC. Overexpression of BRPF1 was (Supplementary Fig. S1), is a major component of the MOZ/ related to a poor survival rate in HCC patients. BRPF1 is an MORF acetyltransferase complex (Fig. 1f). We thus speculated epigenetic reader protein containing a bromodomain, two PHD that activation of the BRPF1/MOZ/MORF complex may play an fingers and a PWWP domain. BRPF1 is mostly known as a important role in liver carcinogenesis. component of the MOZ/MORF acetylation complex15. It has been reported that BRPF1 is critical for the formation of this complex as Clinicopathological relevance of BRPF1 upregulation in it links ING5 and MOZ/MORF together. BRPF1 also plays a role human HCC. BRPF1 has two alternative splicing isoforms in regulating the histone acetyltransferase activity of MOZ/ (BRPF1A and BRPF1B) (Fig. 2a). BRPF1B encodes functional MORF15,16. It has been proposed that the bromodomain of BRPF1 BRPF1 protein, while BRPF1A uses an alternative splice site at may contribute to the chromatin binding and target specificity of exon 6, leading to an insertion of 6 extra amino acids. This the MOZ/MORF complex, wherein the catalytic subunit MOZ/ insertion results in a structural change in the bromodomain that MORF catalyzes H3K9, H3K14, and H3K23 acetylation to activate hinders its binding with acetylated lysine residue27, which causes gene transcription16–18. BRPF1 is indispensable for embryonic BRPF1A to become a functionally inactive isoform. To distin- development. Mouse genetic studies have shown that BRPF1 guish these two BRPF1 isoforms, we reanalyzed our RNA-seq raw knockout affects hematopoiesis as well as brain development and data with an isoform-specific algorithm. We found that BRPF1B even causes early lethality19,20. Moreover, mutation of BRPF1 was the predominant isoform in HCC samples, accounting for causes intellectual disability and facial dysmorphisms in ~80% of the BRPF1 transcripts. This isoform-specific analysis also humans21. Recent studies demonstrated that BRPF1 is recurrently confirmed that only the functional BRPF1B isoform was sig- mutated in adult Shh medulloblastoma22 and also critical for nificantly upregulated in human HCC,
Load more

Recommended publications
  • Functional Roles of Bromodomain Proteins in Cancer
    cancers Review Functional Roles of Bromodomain Proteins in Cancer Samuel P. Boyson 1,2, Cong Gao 3, Kathleen Quinn 2,3, Joseph Boyd 3, Hana Paculova 3 , Seth Frietze 3,4,* and Karen C. Glass 1,2,4,* 1 Department of Pharmaceutical Sciences, Albany College of Pharmacy and Health Sciences, Colchester, VT 05446, USA; [email protected] 2 Department of Pharmacology, Larner College of Medicine, University of Vermont, Burlington, VT 05405, USA; [email protected] 3 Department of Biomedical and Health Sciences, University of Vermont, Burlington, VT 05405, USA; [email protected] (C.G.); [email protected] (J.B.); [email protected] (H.P.) 4 University of Vermont Cancer Center, Burlington, VT 05405, USA * Correspondence: [email protected] (S.F.); [email protected] (K.C.G.) Simple Summary: This review provides an in depth analysis of the role of bromodomain-containing proteins in cancer development. As readers of acetylated lysine on nucleosomal histones, bromod- omain proteins are poised to activate gene expression, and often promote cancer progression. We examined changes in gene expression patterns that are observed in bromodomain-containing proteins and associated with specific cancer types. We also mapped the protein–protein interaction network for the human bromodomain-containing proteins, discuss the cellular roles of these epigenetic regu- lators as part of nine different functional groups, and identify bromodomain-specific mechanisms in cancer development. Lastly, we summarize emerging strategies to target bromodomain proteins in cancer therapy, including those that may be essential for overcoming resistance. Overall, this review provides a timely discussion of the different mechanisms of bromodomain-containing pro- Citation: Boyson, S.P.; Gao, C.; teins in cancer, and an updated assessment of their utility as a therapeutic target for a variety of Quinn, K.; Boyd, J.; Paculova, H.; cancer subtypes.
    [Show full text]
  • Function of Bromodomain and Extra-Terminal Motif Proteins (Bets) in Gata1-Mediated Transcription
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2015 Function of Bromodomain and Extra-Terminal Motif Proteins (bets) in Gata1-Mediated Transcription Aaron James Stonestrom University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Molecular Biology Commons, and the Pharmacology Commons Recommended Citation Stonestrom, Aaron James, "Function of Bromodomain and Extra-Terminal Motif Proteins (bets) in Gata1-Mediated Transcription" (2015). Publicly Accessible Penn Dissertations. 1148. https://repository.upenn.edu/edissertations/1148 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/1148 For more information, please contact [email protected]. Function of Bromodomain and Extra-Terminal Motif Proteins (bets) in Gata1-Mediated Transcription Abstract Bromodomain and Extra-Terminal motif proteins (BETs) associate with acetylated histones and transcription factors. While pharmacologic inhibition of this ubiquitous protein family is an emerging therapeutic approach for neoplastic and inflammatory disease, the mechanisms through which BETs act remain largely uncharacterized. Here we explore the role of BETs in the physiologically relevant context of erythropoiesis driven by the transcription factor GATA1. First, we characterize functions of the BET family as a whole using a pharmacologic approach. We find that BETs are broadly required for GATA1-mediated transcriptional activation, but that repression is largely BET-independent. BETs support activation by facilitating both GATA1 occupancy and transcription downstream of its binding. Second, we test the specific olesr of BETs BRD2, BRD3, and BRD4 in GATA1-activated transcription. BRD2 and BRD4 are required for efficient anscriptionaltr activation by GATA1. Despite co-localizing with the great majority of GATA1 binding sites, we find that BRD3 is not equirr ed for GATA1-mediated transcriptional activation.
    [Show full text]
  • Bromodomain Protein BRDT Directs ΔNp63 Function and Super
    Cell Death & Differentiation (2021) 28:2207–2220 https://doi.org/10.1038/s41418-021-00751-w ARTICLE Bromodomain protein BRDT directs ΔNp63 function and super-enhancer activity in a subset of esophageal squamous cell carcinomas 1 1 2 1 1 2 Xin Wang ● Ana P. Kutschat ● Moyuru Yamada ● Evangelos Prokakis ● Patricia Böttcher ● Koji Tanaka ● 2 3 1,3 Yuichiro Doki ● Feda H. Hamdan ● Steven A. Johnsen Received: 25 August 2020 / Revised: 3 February 2021 / Accepted: 4 February 2021 / Published online: 3 March 2021 © The Author(s) 2021. This article is published with open access Abstract Esophageal squamous cell carcinoma (ESCC) is the predominant subtype of esophageal cancer with a particularly high prevalence in certain geographical regions and a poor prognosis with a 5-year survival rate of 15–25%. Despite numerous studies characterizing the genetic and transcriptomic landscape of ESCC, there are currently no effective targeted therapies. In this study, we used an unbiased screening approach to uncover novel molecular precision oncology targets for ESCC and identified the bromodomain and extraterminal (BET) family member bromodomain testis-specific protein (BRDT) to be 1234567890();,: 1234567890();,: uniquely expressed in a subgroup of ESCC. Experimental studies revealed that BRDT expression promotes migration but is dispensable for cell proliferation. Further mechanistic insight was gained through transcriptome analyses, which revealed that BRDT controls the expression of a subset of ΔNp63 target genes. Epigenome and genome-wide occupancy studies, combined with genome-wide chromatin interaction studies, revealed that BRDT colocalizes and interacts with ΔNp63 to drive a unique transcriptional program and modulate cell phenotype. Our data demonstrate that these genomic regions are enriched for super-enhancers that loop to critical ΔNp63 target genes related to the squamous phenotype such as KRT14, FAT2, and PTHLH.
    [Show full text]
  • Recognition of Cancer Mutations in Histone H3K36 by Epigenetic Writers and Readers Brianna J
    EPIGENETICS https://doi.org/10.1080/15592294.2018.1503491 REVIEW Recognition of cancer mutations in histone H3K36 by epigenetic writers and readers Brianna J. Kleina, Krzysztof Krajewski b, Susana Restrepoa, Peter W. Lewis c, Brian D. Strahlb, and Tatiana G. Kutateladzea aDepartment of Pharmacology, University of Colorado School of Medicine, Aurora, CO, USA; bDepartment of Biochemistry & Biophysics, The University of North Carolina School of Medicine, Chapel Hill, NC, USA; cWisconsin Institute for Discovery, University of Wisconsin, Madison, WI, USA ABSTRACT ARTICLE HISTORY Histone posttranslational modifications control the organization and function of chromatin. In Received 30 May 2018 particular, methylation of lysine 36 in histone H3 (H3K36me) has been shown to mediate gene Revised 1 July 2018 transcription, DNA repair, cell cycle regulation, and pre-mRNA splicing. Notably, mutations at or Accepted 12 July 2018 near this residue have been causally linked to the development of several human cancers. These KEYWORDS observations have helped to illuminate the role of histones themselves in disease and to clarify Histone; H3K36M; cancer; the mechanisms by which they acquire oncogenic properties. This perspective focuses on recent PTM; methylation advances in discovery and characterization of histone H3 mutations that impact H3K36 methyla- tion. We also highlight findings that the common cancer-related substitution of H3K36 to methionine (H3K36M) disturbs functions of not only H3K36me-writing enzymes but also H3K36me-specific readers. The latter case suggests that the oncogenic effects could also be linked to the inability of readers to engage H3K36M. Introduction from yeast to humans and has been shown to have a variety of functions that range from the control Histone proteins are main components of the of gene transcription and DNA repair, to cell cycle nucleosome, the fundamental building block of regulation and nutrient stress response [8].
    [Show full text]
  • Further Delineation of the Clinical Spectrum of KAT6B Disorders and Allelic Series of Pathogenic Variants
    ARTICLE © American College of Medical Genetics and Genomics Further delineation of the clinical spectrum of KAT6B disorders and allelic series of pathogenic variants Li Xin Zhang1, Gabrielle Lemire, MD2, Claudia Gonzaga-Jauregui, PhD3, Sirinart Molidperee, Gr. Cert1, Carolina Galaz-Montoya, MD3, David S. Liu, MD3, Alain Verloes, MD PhD4, Amelle G. Shillington, DO5, Kosuke Izumi, MD PhD6, Alyssa L. Ritter, MS LCGC7, Beth Keena, MS LCGC6, Elaine Zackai, MD7,8, Dong Li, PhD9, Elizabeth Bhoj, MD PhD7, Jennifer M. Tarpinian, MS CGC10, Emma Bedoukian, MS LCGC10, Mary K. Kukolich, MD11, A. Micheil Innes, MD12, Grace U. Ediae, BSc13, Sarah L. Sawyer, MD PhD14, Karippoth Mohandas Nair, MD15, Para Chottil Soumya, MD15, Kinattinkara R. Subbaraman, MD15, Frank J. Probst, MD PhD3,16, Jennifer A. Bassetti, MD3,16, Reid V. Sutton, MD3,16, Richard A. Gibbs, PhD3, Chester Brown, MD PhD17, Philip M. Boone, MD PhD18, Ingrid A. Holm, MD MPH18, Marco Tartaglia, PhD19, Giovanni Battista Ferrero, MD PhD20, Marcello Niceta, MD PhD19, Maria Lisa Dentici, MD19, Francesca Clementina Radio, MD PhD19, Boris Keren, MD21, Constance F. Wells, MD22, Christine Coubes, MD22, Annie Laquerrière, MD PhD23, Jacqueline Aziza, MD24, Charlotte Dubucs, MD24, Sheela Nampoothiri, MD25, David Mowat, MD26, Millan S. Patel, MD27, Ana Bracho, MD28, Francisco Cammarata-Scalisi, MD29, Alper Gezdirici, MD30, Alberto Fernandez-Jaen, MD31, Natalie Hauser, MD32, Yuri A. Zarate, MD33, Katherine A. Bosanko, MS CGC33, Klaus Dieterich, MD PhD34, John C. Carey, MD MPH35, Jessica X. Chong, PhD36,37, Deborah A. Nickerson, PhD37,38, Michael J. Bamshad, MD36,37, Brendan H. Lee, MD PhD3, Xiang-Jiao Yang, PhD39, James R.
    [Show full text]
  • Download Product Insert (PDF)
    PRODUCT INFORMATION BRD3 bromodomains 1 and 2 (human, recombinant) Item No. 14864 Overview and Properties Synonyms: Bromodomain containing protein 3, ORFX, RING3L, RING3-like protein Source: Recombinant N-terminal GST-tagged protein expressed in E. coli Amino Acids: 2-434 Uniprot No.: Q15059 Molecular Weight: 75.0 kDa Storage: -80°C (as supplied) Stability: ≥2 years Purity: batch specific (≥85% estimated by SDS-PAGE) Supplied in: 50 mM Tris, pH 8.0, with 150 mM sodium chloride and 20% glycerol Protein Concentration: batch specific mg/ml Activity: batch specific U/ml Specific Activity: batch specific U/mg Information represents the product specifications. Batch specific analytical results are provided on each certificate of analysis. Image 1 2 3 4 · · · · · · ·250 kDa · · · · · · ·150 kDa · · · · · · ·100 kDa · · · · · · ·75 kDa · · · · · · ·50 kDa · · · · · · ·37 kDa · · · · · · ·25 kDa · · · · · · ·20 kDa · · · · · · ·15 kDa Lane 1: BRD3 (4 µg) Lane 2: BRD3 (6 µg) Lane 3: BRD3 (8 µg) Lane 4: MW Markers WARNING CAYMAN CHEMICAL THIS PRODUCT IS FOR RESEARCH ONLY - NOT FOR HUMAN OR VETERINARY DIAGNOSTIC OR THERAPEUTIC USE. 1180 EAST ELLSWORTH RD SAFETY DATA ANN ARBOR, MI 48108 · USA This material should be considered hazardous until further information becomes available. Do not ingest, inhale, get in eyes, on skin, or on clothing. Wash thoroughly after handling. Before use, the user must review the complete Safety Data Sheet, which has been sent via email to your institution. PHONE: [800] 364-9897 WARRANTY AND LIMITATION OF REMEDY [734] 971-3335 Buyer agrees to purchase the material subject to Cayman’s Terms and Conditions. Complete Terms and Conditions including Warranty and Limitation of Liability information can be found on our website.
    [Show full text]
  • Epigenetic Reader BRD4 Inhibition As a Therapeutic Strategy to Suppress E2F2-Cell Cycle Regulation Circuit in Liver Cancer
    www.impactjournals.com/oncotarget/ Oncotarget, Vol. 7, No. 22 Epigenetic reader BRD4 inhibition as a therapeutic strategy to suppress E2F2-cell cycle regulation circuit in liver cancer Seong Hwi Hong1,*, Jung Woo Eun2,*, Sung Kyung Choi1, Qingyu Shen2, Wahn Soo Choi1, Jeung-Whan Han3, Suk Woo Nam2, Jueng Soo You1 1Konkuk University Medical Centre, School of Medicine, Konkuk University, Seoul 143-701, Korea 2Functional RNomics Research Center, College of Medicine, The Catholic University, Seoul 137-701, Korea 3Research Center for Epigenome Regulation, School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Korea *These authors have contributed equally to this work Correspondence to: Suk Woo Nam, email: [email protected] Jueng Soo You, email: [email protected] Keywords: BET protein, epigenetic components, JQ1, hepatocellular carcinoma (HCC), E2F Received: January 07, 2016 Accepted: March 28, 2016 Published: April 12, 2016 ABSTRACT Deregulation of the epigenome component affects multiple pathways in the cancer phenotype since the epigenome acts at the pinnacle of the hierarchy of gene expression. Pioneering work over the past decades has highlighted that targeting enzymes or proteins involved in the epigenetic regulation is a valuable approach to cancer therapy. Very recent results demonstrated that inhibiting the epigenetic reader BRD4 has notable efficacy in diverse cancer types. We investigated the potential of BRD4 as a therapeutic target in liver malignancy. BRD4 was overexpressed in three different large cohort of hepatocellular carcinoma (HCC) patients as well as in liver cancer cell lines. BRD4 inhibition by JQ1 induced anti-tumorigenic effects including cell cycle arrest, cellular senescence, reduced wound healing capacity and soft agar colony formation in liver cancer cell lines.
    [Show full text]
  • Recombinant BRPF1 (627-746) Protein Catalog No: 31375, 31875 Quantity: 100, 1000 Μg Lot No: 32717002 Concentration: 1.3 Μg/Μl Expressed In: E
    Recombinant BRPF1 (627-746) protein Catalog No: 31375, 31875 Quantity: 100, 1000 µg Lot No: 32717002 Concentration: 1.3 µg/µl Expressed In: E. coli Source: Human Buffer Contents: Recombinant BRPF1 (627-746) protein expressed in E. coli and provided in 25 mM Tris pH 8, 300 mM NaCl and 10% glycerol. Background: Bromodomain and PHD finger containing (BRPF1) protein is a component of the MOZ/MORF complex which has histone H3 acetyltransferase activity. The bromodomain of BRPF1 recognizes acetylated histone lysine residues and function as a ‘reader’ of these epigenetic histone marks to regulate chromatin structure and gene expression by linking associated proteins to the recognized acetylated nucleosomal targets. BRPF1 is known to positively regulate the transcription of RUNX1 and RUNX2. Protein Details: The peptide corresponding to amino acids 627 - 746 that contains the bromodomain sequences of BRPF1 (accession number NM_001003694.1) was expressed in E. coli and contains an N-terminal His-Tag and C- terminal FLAG-Tag with an observed molecular weight of 20.3 kDa. The recombinant protein is >90% pure by SDS- PAGE. Application Notes: Recombinant BRPF1 (627-746) is suitable for use in binding assays, inhibitor screening, and selectivity profiling. Storage and Guarantee: Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage. This product is guaranteed for 6 months from date of receipt. This product is for research use only and is not for use in diagnostic procedures. North America 877 222 9543 • Europe +32 (0)2 653 0001 • Japan +81 (0)3 5225 3638 • www.activemotif.com Recombinant BRPF1 (627-746) protein gel.
    [Show full text]
  • BRD2-2 (His) (Bromodomain Containing Protein 2 (RING3), Bromodomain 2)
    BRD2-2 (His) (Bromodomain containing protein 2 (RING3), bromodomain 2) CATALOG NO.: RD-11-145 LOT NO.: DESCRIPTION: Human recombinant BRD2, bromodomain-2 (residues 344-454; Genbank Accession # NM_005104; MW = 15.7 kDa) expressed in E. coli with an N-terminal His-tag. BRD2, like other human members of the BET family of chromatin-binding proteins (BRD3, BRD4, BRDT), comprises two bromodomains (see reviews 1,2 ), protein modules that bind ε-N-acetyllysine residues 3,4 . When overexpressed in 293 cells, BRD2, along with BRD3, binds the hyperacetylated chromatin of transcribed genes, regions enriched in acetylated histone H4 lysine-5 (H4K5Ac), H4K12Ac, H3K14Ac, but deficient in H4K16Ac and H3K9me 5. A single H4K5AcK12Ac peptide can bind two copies of BRD2-2 (BRD2, bromodomain 2), each interacting with one of the two acetylated lysines 6. In an in vitro RNA polymerase II transcription system, binding of either BRD2 or BRD3 to a chromatin template assembled with hyperacetylated histones enabled transcription through the nucleosomes 5. Further, BRD2 displayed histone chaperone activity, catalyzing the transfer of histone octamers from hyperacetylated oligonucleosomes to a labeled 190 bp 5s rDNA fragment 5. Like BRD4, BRD2 is a ubiquitously expressed 7 transcriptional regulator 8 and atypical protein kinase 9, with functions in cell cycle progression 8 and embryogenesis 10,11 . BRD2 binds preferentially to hyperacetylated histone H4 in H2A.Z-containing nucleosomes and this interaction is required for activation of androgen receptor (AR)-regulated genes in prostate cancer cells 12 . In addition to prostate cancer, leukemia is a potential indication for specific BRD2 inhibition9,13 . BRD2 suppresses HIV transcription in latently infected cells and may therefore represent a target in therapeutic strategies involving viral reactivation 14 .
    [Show full text]
  • BRD2 Bromodomains 1 and 2 TR-FRET Assay Kit
    BRD2 bromodomains 1 and 2 TR-FRET Assay Kit Item No. 600810 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd · Ann Arbor, MI · USA TABLE OF CONTENTS GENERAL INFORMATION GENERAL INFORMATION 3 Materials Supplied Materials Supplied 4 Safey Data 4 Precautions Kit will arrive packaged as a -80°C kit. For best results, remove components and store as stated below. 5 If You Have Problems 5 Storage and Stability Item Item 384 wells 1,920 wells 9,600 wells Storage 5 Materials Needed but Not Supplied Number Quantity/Size Quantity/ Quantity/Size Size INTRODUCTION 6 Background 600811 BRD2 bromodomains 1 vial/420 wells 5 vials/ 5 vials/ -80°C 7 About This Assay 1 and 2 Europium 420 wells 2,100 wells Chelate 8 Introduction to TR-FRET 600812 BRD2 bromodomains 1 vial/420 wells 5 vials/ 5 vials/ -80°C PRE-ASSAY PREPARATION 11 Buffer Preparation 1 and 2 Ligand/APC 420 wells 2,100 wells Acceptor Mixture 11 Sample Preparation 600503 TR-FRET Assay 1 vial/2 ml 1 vial/10 ml 5 vials/10 ml -20°C ASSAY PROTOCOL 12 Preparation of Assay-Specific Reagents Buffer (10X) 14 Performing the Assay 600504 TR-FRET Assay 1 vial/200 mg 1 vial/1 g 5 vials/1 g -20°C 16 Effects of Solvent Buffer Additive ANALYSIS 17 Calculations 600505 (+)-JQ1 Positive 1 vial/8 nmol 5 vials/8 nmol 5 vials/40 nmol -80°C Control 17 Performance Characteristics 400093 384-Well Solid Plate 1 plate 5 plates 25 plates RT RESOURCES 20 Troubleshooting (low volume; black) 21 References 400023 Foil Plate Covers 1 cover 5 covers 25 covers RT 23 Notes 23 Warranty and Limitation of Remedy If any of the items listed above are damaged or missing, please contact our Customer Service department at (800) 364-9897 or (734) 971-3335.
    [Show full text]
  • Deficiency of Intellectual Disability-Related Gene Brpf1 Reduced Inhibitory
    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.11.426233; this version posted January 12, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Deficiency of intellectual disability-related gene Brpf1 reduced inhibitory 2 neurotransmission and Map2k7 expression in GABAergic interneurons 3 Jingli Cao1, Weiwei Xian1, Maierdan Palihati1, Yu Zhu1, Guoxiang Wang2, Yunli Xie2, 4 Guomin Zhou1,3#, Linya You1# 5 6 1Department of Human Anatomy & Histoembryology, School of Basic Medical 7 Sciences, Fudan University, Shanghai 200032, China 8 2Institutes of Brain Sciences, Fudan University, Shanghai 200032, China 9 3Key Laboratory of Medical Imaging Computing and Computer Assisted Intervention 10 of Shanghai, Shanghai 200032, China 11 12 #Correspondences to: [email protected]; [email protected]. 13 14 Conflict of interest: The authors declare no conflicts of interest. 15 16 Running Title: Brpf1 loss reduced neurotransmission 17 18 Keywords: intellectual disability; Brpf1; MGE-derived GABAergic interneurons; 19 differentiation; dendritic morphology, electrophysiology; in vivo migration; 20 mRNA-Seq 21 22 bioRxiv preprint doi: https://doi.org/10.1101/2021.01.11.426233; this version posted January 12, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 23 Abstract 24 Intellectual disability is closely related to impaired GABA neurotransmission.
    [Show full text]
  • Histone Acetyltransferase KAT6A Upregulates PI3K/AKT Signaling Through TRIM24 Binding Deguan Lv1,2, Feng Jia3, Yanli Hou4, Youzhou Sang2, Angel A
    Published OnlineFirst October 11, 2017; DOI: 10.1158/0008-5472.CAN-17-1388 Cancer Tumor and Stem Cell Biology Research Histone Acetyltransferase KAT6A Upregulates PI3K/AKT Signaling through TRIM24 Binding Deguan Lv1,2, Feng Jia3, Yanli Hou4, Youzhou Sang2, Angel A. Alvarez5, Weiwei Zhang2, Wei-Qiang Gao2,BoHu5, Shi-Yuan Cheng2,5, Jianwei Ge3, Yanxin Li6, and Haizhong Feng2 Abstract Lysine acetyltransferase KAT6A is a chromatin regulator that PI3K/AKT signaling and tumorigenesis. Overexpressing activated contributes to histone modification and cancer, but the basis of its AKT or PIK3CA rescued the growth inhibition due to KAT6A actions are not well understood. Here, we identify a KAT6A silencing. Conversely, the pan-PI3K inhibitor LY294002 abrogat- signaling pathway that facilitates glioblastoma (GBM), where it ed the growth-promoting effect of KAT6A. Overexpression is upregulated. KAT6A expression was associated with GBM of KAT6A or TRIM24, but not KAT6A acetyltransferase activity– patient survival. KAT6A silencing suppressed cell proliferation, deficient mutants or TRIM24 mutants lacking H3K23ac-bind- cell migration, colony formation, and tumor development in an ing sites, promoted PIK3CA expression, AKT phosphorylation, orthotopic mouse xenograft model system. Mechanistic investi- and cell proliferation. Taken together, our results define gations demonstrated that KAT6A acetylates lysine 23 of histone an essential role of KAT6A in glioma formation, rationalizing H3 (H3K23), which recruits the nuclear receptor binding protein its candidacy as a therapeutic target for GBM treatment. TRIM24 to activate PIK3CA transcription, thereby enhancing Cancer Res; 77(22); 6190–201. Ó2017 AACR. Introduction deregulated in cancer, including glioma, partially due to aberrant histone modifications and mutations (2–4).
    [Show full text]