DOCSLIB.ORG

Mutational Landscape and Antiproliferative Functions of ELF

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mutational Landscape and Antiproliferative Functions of ELF Published OnlineFirst February 26, 2016; DOI: 10.1158/0008-5472.CAN-14-3816 Cancer Molecular and Cellular Pathobiology Research Mutational Landscape and Antiproliferative Functions of ELF Transcription Factors in Human Cancer Mizuo Ando1,2, Masahito Kawazu3, Toshihide Ueno1, Daizo Koinuma4, Koji Ando5, Junji Koya6, Keisuke Kataoka6, Takahiko Yasuda1, Hiroyuki Yamaguchi1, Kazutaka Fukumura1, Azusa Yamato1, Manabu Soda1, Eirin Sai3, Yoshihiro Yamashita1, Takahiro Asakage2, Yasushi Miyazaki5, Mineo Kurokawa6, Kohei Miyazono4, Stephen D. Nimer7, Tatsuya Yamasoba2, and Hiroyuki Mano1,8 Abstract ELF4 (also known as MEF) is a member of the ETS family of ELF1 and ELF2, as in ELF4, were widespread across human transcription factors. An oncogenic role for ELF4 has been cancers, but were almost all mutually exclusive. Moreover, demonstrated in hematopoietic malignancies, but its function chromatin immunoprecipitation coupled with high-throughput in epithelial tumors remains unclear. Here, we show that ELF4 sequencing revealed ELF4-binding sites in genomic regions can function as a tumor suppressor and is somatically inacti- adjacent to genes related to cell-cycle regulation and apoptosis. vated in a wide range of human tumors. We identified a Finally, we provide mechanistic evidence that the antiprolifera- missense mutation affecting the transactivation potential of tive effects of ELF4 were mediated through the induction of ELF4 in oral squamous cell carcinoma cells. Restoration of the HRK, an activator of apoptosis, and DLX3, an inhibitor of cell transactivation activity through introduction of wild-type ELF4 growth. Collectively, our findings reveal a novel subtype of significantly inhibited cell proliferation in vitro and tumor human cancer characterized by inactivating mutations in the xenograft growth. Furthermore, we found that ELF1 and ELF2, ELF subfamily of proteins, and warrant further investigation of closely related transcription factors to ELF4, also exerted anti- the specific settings where ELF restoration may be therapeuti- proliferative effects in multiple cancer cell lines. Mutations in cally beneficial. Cancer Res; 76(7); 1–11. Ó2016 AACR. Introduction prostate tumors (3). Forced expression of TMPRSS2-ERG in the context of PI3K pathway activation in prostate epithelial cells ELF4, also known as myeloid Elf-1–like factor (MEF), belongs results in the formation of prostatic intraepithelial neoplasia in to the ETS family of transcription factors that are encoded by 28 mice, demonstrating an oncogenic role for the fusion gene (4). In independent genes in the human genome (1, 2). ETS proteins are addition, fusion of EWSR1 with the ETS family gene FLI1 or implicated in the proliferation, differentiation, and malignant related ones occurs frequently in Ewing's sarcoma (5). Such transformation of various cell lineages. The genes for ERG or fusions are promising therapeutic targets, given that knockdown members of the PEA3 subfamily (ETV1, ETV4, and ETV5) of ETS of EWSR1–FLI1 expression suppresses sarcoma growth. proteins, for instance, are fused to TMPRSS2 in >50% of human ELF4 as well as ELF1 and ELF2 constitute the ELF subfamily of ETS proteins. Elf4-null mice manifest severe defects in the devel- 1Department of Cellular Signaling, Graduate School of Medicine, The opment and function of natural killer cells and NK-T cells (6). University of Tokyo, Tokyo, Japan. 2Department of Otolaryngology Furthermore, the hematopoietic system of such mice harbors an and Head and Neck Surgery, Graduate School of Medicine, The Uni- increased population of quiescent hematopoietic stem cells com- versity of Tokyo, Tokyo, Japan. 3Department of Medical Genomics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. pared with wild-type animals, suggesting that ELF4 plays a key 4 Department of Molecular Pathology, Graduate School of Medicine, role in the transition from G0 to G1 phases of the cell cycle (7). The 5 The University of Tokyo, Tokyo, Japan. Department of Hematology, activity of ELF4 is selectively increased in G1 phase and is sup- Nagasaki University Graduate School of Biomedical Sciences, Naga- pressed by the complex of cyclin A and cyclin-dependent kinase 2 saki, Japan. 6Department of Hematology and Oncology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. 7Sylvester (CDK2), further linking ELF4 to cell-cycle regulation (8). Comprehensive Cancer Center, University of Miami, Miami, Florida. An oncogenic role for ELF4 has been suggested by several 8 Strategic Basic Research Program, Japan Science and Technology observations. ELF4 is highly expressed in clinical specimens of Agency, Saitama, Japan. leukemia and ovarian cancer (9,10). Elf4 induces Mdm2, and also Note: Supplementary data for this article are available at Cancer Research inhibits Cdkn2a activation, and thereby cooperates with onco- Online (http://cancerres.aacrjournals.org/). genic HRAS(G12V) in malignant transformation (11). Further- Corresponding Author: Hiroyuki Mano, The University of Tokyo, 7-3-1 Hongo, more, cells from Elf4-null mice are impaired in the ability to form Bunkyo-ku, Tokyo 113-0033, Japan. Phone: 81-3-5841-0633; Fax: 81-3-5841- neurospheres, and knockdown of ELF4 was found to inhibit 0634; E-mail: [email protected] neurosphere formation by human primary glioma stem-like cells, doi: 10.1158/0008-5472.CAN-14-3816 suggesting that ELF4 promotes stemness in glioma cells (12). On Ó2016 American Association for Cancer Research. the other hand, however, one article shows a possibility for the www.aacrjournals.org OF1 Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 2016 American Association for Cancer Research. Published OnlineFirst February 26, 2016; DOI: 10.1158/0008-5472.CAN-14-3816 Ando et al. growth-suppressive role of ELF4 in a human lung cancer cell line resulted in the identification of 12 missense mutations including A549 (13). ELF4(L211M) (20). Given that L211 maps within the conserved We now describe an L211M amino acid substitution in the ETS ETS domain of ELF4 (Fig. 1A), we chose this nonsynonymous domain of ELF4 that is present in the human squamous cell mutation for further analysis. A homozygous V382I mutation of carcinoma cell line T3M-1 Cl-10 (14). Forced expression of wild- ELF4 was also identified in the genome of the human breast type ELF4 resulted in marked inhibition of cell proliferation not cancer cell line MDA-MB-231 (data not shown). only in this cell line but also in a human fibrosarcoma cell line Further searching for nonsynonymous mutations of ELF4 in HT1080 (15) and a human endometrial carcinoma cell line the Catalogue of Somatic Mutations in Cancer (COSMIC; http:// HEC59 (16) as well as in the nontransformed human mammary cancer.sanger.ac.uk/cancergenome/projects/cosmic), Cancer Cell epithelial cell line MCF10A (17). We further demonstrate that Line Encyclopedia (CCLE; http://www.broadinstitute.org/ccle/ somatic mutations of ELF1, ELF2, and ELF4 are present and home), International Cancer Genome Consortium (ICGC; mutually exclusive in a wide range of human tumors, and that http://icgc.org), and The Cancer Genome Atlas (TCGA; https:// the antiproliferative effect of ELF4 is mediated through the induc- tcga-data.nci.nih.gov/tcga/tcgaHome2.jsp) databases revealed tion of HRK and DLX3 expression. >40 mutations widely distributed throughout the protein struc- ture of ELF4 (Fig. 1A). In addition, Totoki and colleagues (21) fi BCORL1–ELF4 Materials and Methods identi ed a fusion gene in hepatocellular carcinoma. Cell lines We examined the transactivation potential of ELF4(L211M), T3M-1 Cl-10 or HEC59 was obtained from RIKEN Cell Bank or ELF4(V382I), and BCORL1-ELF4 in comparison with that of the Japanese Collection of Research Bioresources Cell Bank, respec- wild-type protein (Fig. 1B). The activity of a reporter construct tively, and human embryonic kidney 293T (HEK293T), HT1080, containing the ETS-binding regions within the IL3 promoter MDA-MB-231, KATO-III, A549 and MCF10A were obtained (APET; ref. 19) was markedly increased by expression of wild- from ATCC. MCF10A was maintained in DMEM-F12 supplemen- type ELF4. Whereas the V382I substitution did not substantially ted with 5% horse serum (Biowest), recombinant human EGF affect the transactivation activity of ELF4, the activities of both (20 ng/mL; Peprotech), bovine insulin (10 mg/mL; Sigma- ELF4(L211M) and BCORL1-ELF4 were significantly attenuated Aldrich), hydrocortisone (0.5 mg/mL; Sigma-Aldrich), and cholera compared with that of the wild-type protein. The observed tran- toxin (100 ng/mL; Sigma-Aldrich). All other cells were main- scriptional activity was dependent on the ETS-binding regions of tained in DMEM-F12 supplemented with 10% FBS and 2 mmol/L the IL3 promoter, given that substitution of nucleotides within the L-glutamine (Invitrogen). ETS-binding sites (the mAPET reporter) abolished all activity. Furthermore, expression level of endogenous as well as artificially Functional analysis of ELF4 and other proteins introduced ELF4 mRNA was determined by reverse transcription We inserted an internal ribosomal entry site (ires) and (RT) and real-time PCR (PCR) analysis. As demonstrated in enhanced green fluorescent protein (EGFP) cDNA (Clontech) Supplementary Fig. S1, almost 20 times larger amounts of ELF4 into pMXS (18) to yield the pMXS-ires-EGFP plasmid, which mRNA were expressed in the transfected cells compared with the allows simultaneous expression of inserted cDNAs together with control, suggesting that competitive effects of endogenous ELF4 EGFP. For reporter assays of ELF4 activity, HEK293T cells were are negligible. transfected with pMXS- or pMXS-ires-EGFP-based expression We next examined whether such loss of function of ELF4 plasmids for various cDNAs, the firefly luciferase–based APET might contribute to carcinogenesis. Whereas introduction of reporter plasmid (19), and the pGL-TK plasmid for Renilla lucif- ELF4(L211M) or BCORL1-ELF4 had no marked effect on the erase (Promega). As a negative control for ELF4 activity, cells were proliferation of T3M-1 Cl-10 cells, expression of wild-type ELF4 transfected with the mAPET plasmid (19) instead of APET. Quan- or ELF4(V382I) greatly inhibited cell growth (Fig.
Load more

Recommended publications
  • Targeting BCL-2 in Cancer: Advances, Challenges, and Perspectives
    cancers Review Targeting BCL-2 in Cancer: Advances, Challenges, and Perspectives Shirin Hafezi 1 and Mohamed Rahmani 1,2,* 1 Research Institute of Medical & Health Sciences, University of Sharjah, P.O. Box 27272 Sharjah, United Arab Emirates; [email protected] 2 Department of Basic Medical Sciences, College of Medicine, University of Sharjah, P.O. Box 27272 Sharjah, United Arab Emirates * Correspondence: [email protected]; Tel.: +971-6505-7759 Simple Summary: Apoptosis, a programmed form of cell death, represents the main mechanism by which cells die. Such phenomenon is highly regulated by the BCL-2 family of proteins, which includes both pro-apoptotic and pro-survival proteins. The decision whether cells live or die is tightly controlled by a balance between these two classes of proteins. Notably, the pro-survival Bcl-2 proteins are frequently overexpressed in cancer cells dysregulating this balance in favor of survival and also rendering cells more resistant to therapeutic interventions. In this review, we outlined the most important steps in the development of targeting the BCL-2 survival proteins, which laid the ground for the discovery and the development of the selective BCL-2 inhibitor venetoclax as a therapeutic drug in hematological malignancies. The limitations and future directions are also discussed. Abstract: The major form of cell death in normal as well as malignant cells is apoptosis, which is a programmed process highly regulated by the BCL-2 family of proteins. This includes the antiapoptotic proteins (BCL-2, BCL-XL, MCL-1, BCLW, and BFL-1) and the proapoptotic proteins, which can be divided into two groups: the effectors (BAX, BAK, and BOK) and the BH3-only proteins Citation: Hafezi, S.; Rahmani, M.
    [Show full text]
  • Multivariate Meta-Analysis of Differential Principal Components Underlying Human Primed and Naive-Like Pluripotent States
    bioRxiv preprint doi: https://doi.org/10.1101/2020.10.20.347666; this version posted October 21, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license. October 20, 2020 To: bioRxiv Multivariate Meta-Analysis of Differential Principal Components underlying Human Primed and Naive-like Pluripotent States Kory R. Johnson1*, Barbara S. Mallon2, Yang C. Fann1, and Kevin G. Chen2*, 1Intramural IT and Bioinformatics Program, 2NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA Keywords: human pluripotent stem cells; naive pluripotency, meta-analysis, principal component analysis, t-SNE, consensus clustering *Correspondence to: Dr. Kory R. Johnson ([email protected]) Dr. Kevin G. Chen ([email protected]) 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.10.20.347666; this version posted October 21, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license. ABSTRACT The ground or naive pluripotent state of human pluripotent stem cells (hPSCs), which was initially established in mouse embryonic stem cells (mESCs), is an emerging and tentative concept. To verify this important concept in hPSCs, we performed a multivariate meta-analysis of major hPSC datasets via the combined analytic powers of percentile normalization, principal component analysis (PCA), t-distributed stochastic neighbor embedding (t-SNE), and SC3 consensus clustering.
    [Show full text]
  • ELF2 Antibody - N-Terminal Region Rabbit Polyclonal Antibody Catalog # AI10026
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 ELF2 antibody - N-terminal region Rabbit Polyclonal Antibody Catalog # AI10026 Specification ELF2 antibody - N-terminal region - Product Information Application WB Primary Accession Q15723 Other Accession Q15723-3, NP_006865, NM_006874 Reactivity Human, Mouse, Rat, Rabbit, Dog, Guinea Pig, Horse, Bovine Predicted Human, Mouse, ELF2 antibody - N-terminal region (AI10026) Rat, Rabbit, in Hum. Fetal Heart cells using Western Blot Chicken, Dog, Host: Rabbit Guinea Pig, Horse, Bovine Target Name: ELF2 Host Rabbit Sample Tissue: Human Fetal Heart Clonality Polyclonal Antibody Dilution: 1.0μg/ml Calculated MW 57 kDa KDa ELF2 antibody - N-terminal region - Additional Information Gene ID 1998 Alias Symbol EU32, NERF, NERF-2, NERF-1A, NERF-1B, NERF-1a,b Other Names ETS-related transcription factor Elf-2, ELF2 antibody - N-terminal region (AI10026) E74-like factor 2, New ETS-related factor, in Hum. Fetal Lung cells using Western Blot ELF2 {ECO:0000312|EMBL:AAF671951} Host: Rabbit Target Name: ELF2 Target/Specificity Sample Tissue: Human Fetal Lung The ELF2 gene encodes a protein that Antibody Dilution: 1.0μg/ml physically interacts with AML1 and mediates opposing effects on AML1-mediated transcription of the B cell-specific blk gene. Format Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose. Page 1/2 10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 Reconstitution & Storage Add 50 ul of distilled water. Final anti-ELF2 antibody concentration is 1 mg/ml in PBS buffer with 2% sucrose.
    [Show full text]
  • Dynamics of the BH3-Only Protein Binding Interface of Bcl-Xl
    Biophysical Journal Volume 109 September 2015 1049–1057 1049 Article Dynamics of the BH3-Only Protein Binding Interface of Bcl-xL Xiaorong Liu,1 Alex Beugelsdijk,1 and Jianhan Chen1,* 1Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, Kansas ABSTRACT The balance and interplay between pro-death and pro-survival members of the B-cell lymphoma-2 (Bcl-2) family proteins play key roles in regulation of the mitochondrial pathway of programmed cell death. Recent NMR and biochemical studies have revealed that binding of the proapoptotic BH3-only protein PUMA induces significant unfolding of antiapoptotic Bcl-xL at the interface, which in turn disrupts the Bcl-xL/p53 interaction to activate apoptosis. However, the molecular mecha- nism of such regulated unfolding of Bcl-xL is not fully understood. Analysis of the existing Protein Data Bank structures of Bcl-xL in both bound and unbound states reveal substantial intrinsic heterogeneity at its BH3-only protein binding interface. Large-scale atomistic simulations were performed in explicit solvent for six representative structures to further investigate the intrinsic confor- mational dynamics of Bcl-xL. The results support that the BH3-only protein binding interface of Bcl-xL is much more dynamic compared to the rest of the protein, both unbound and when bound to various BH3-only proteins. Such intrinsic interfacial confor- mational dynamics likely provides a physical basis that allows Bcl-xL to respond sensitively to detailed biophysical properties of the ligand. The ability of Bcl-xL to retain or even enhance dynamics at the interface in bound states could further facilitate the regulation of its interactions with various BH3-only proteins such as through posttranslational modifications.
    [Show full text]
  • Analysis of the Indacaterol-Regulated Transcriptome in Human Airway
    Supplemental material to this article can be found at: http://jpet.aspetjournals.org/content/suppl/2018/04/13/jpet.118.249292.DC1 1521-0103/366/1/220–236$35.00 https://doi.org/10.1124/jpet.118.249292 THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS J Pharmacol Exp Ther 366:220–236, July 2018 Copyright ª 2018 by The American Society for Pharmacology and Experimental Therapeutics Analysis of the Indacaterol-Regulated Transcriptome in Human Airway Epithelial Cells Implicates Gene Expression Changes in the s Adverse and Therapeutic Effects of b2-Adrenoceptor Agonists Dong Yan, Omar Hamed, Taruna Joshi,1 Mahmoud M. Mostafa, Kyla C. Jamieson, Radhika Joshi, Robert Newton, and Mark A. Giembycz Departments of Physiology and Pharmacology (D.Y., O.H., T.J., K.C.J., R.J., M.A.G.) and Cell Biology and Anatomy (M.M.M., R.N.), Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada Received March 22, 2018; accepted April 11, 2018 Downloaded from ABSTRACT The contribution of gene expression changes to the adverse and activity, and positive regulation of neutrophil chemotaxis. The therapeutic effects of b2-adrenoceptor agonists in asthma was general enriched GO term extracellular space was also associ- investigated using human airway epithelial cells as a therapeu- ated with indacaterol-induced genes, and many of those, in- tically relevant target. Operational model-fitting established that cluding CRISPLD2, DMBT1, GAS1, and SOCS3, have putative jpet.aspetjournals.org the long-acting b2-adrenoceptor agonists (LABA) indacaterol, anti-inflammatory, antibacterial, and/or antiviral activity. Numer- salmeterol, formoterol, and picumeterol were full agonists on ous indacaterol-regulated genes were also induced or repressed BEAS-2B cells transfected with a cAMP-response element in BEAS-2B cells and human primary bronchial epithelial cells by reporter but differed in efficacy (indacaterol $ formoterol .
    [Show full text]
  • Adaptive History of the Chimpanzee Subspecies in the Genomic Era
    Adaptive History of the Chimpanzee Subspecies in the Genomic Era Jessica Nye TESI DOCTORAL UPF 2018 Directors de la Tesi Jaume Bertranpetit i Hafid Laayouni CIÈNCIES EXPERIMENTALS I DE LA SALUT ii Acknowledgements I would like to thank my advisors Dr. Jaume Bertranpetit and Dr. Hafid Laayouni for guiding me during my doctoral research. I would like to thank my family and friends for supporting me during my many years of education. To my fellow lab mates, I am grateful for the theoretical discussions we had over the last four years. This work was supported by FI Agaur grant FI-DGR 2015. iii iv Abstract Chimpanzees are our closest living genetic cousins. The species consists of four subspecies, each with a unique demographic history. In order to better understand their species, a detailed map of signatures of selection will highlight important genetic differences between the four subspecies. Here, we interrogate the genome using 15 tests of selection. We simulate neutral and selective scenarios taking their unique demographic history into account in the model. We combine all these elements using a machine learning approach to highlight regions of interest in each subspecies. We specifically investigate the regions of the genome that have been selected after introgression with bonobo. We investigate the haplotype changes that have occurred since the divergence with humans in a few specially selected genes. From this, we find that the evolutionary history of each of the four subspecies is unique. That subtle differences in their demographic history and environment have greatly shaped their genetic diversity. In general, we observe signatures in selection in phenotypes involving immunity, muscle function, reproduction, and DNA repair.
    [Show full text]
  • Original Article a Database and Functional Annotation of NF-Κb Target Genes
    Int J Clin Exp Med 2016;9(5):7986-7995 www.ijcem.com /ISSN:1940-5901/IJCEM0019172 Original Article A database and functional annotation of NF-κB target genes Yang Yang, Jian Wu, Jinke Wang The State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, People’s Republic of China Received November 4, 2015; Accepted February 10, 2016; Epub May 15, 2016; Published May 30, 2016 Abstract: Backgrounds: The previous studies show that the transcription factor NF-κB always be induced by many inducers, and can regulate the expressions of many genes. The aim of the present study is to explore the database and functional annotation of NF-κB target genes. Methods: In this study, we manually collected the most complete listing of all NF-κB target genes identified to date, including the NF-κB microRNA target genes and built the database of NF-κB target genes with the detailed information of each target gene and annotated it by DAVID tools. Results: The NF-κB target genes database was established (http://tfdb.seu.edu.cn/nfkb/). The collected data confirmed that NF-κB maintains multitudinous biological functions and possesses the considerable complexity and diversity in regulation the expression of corresponding target genes set. The data showed that the NF-κB was a central regula- tor of the stress response, immune response and cellular metabolic processes. NF-κB involved in bone disease, immunological disease and cardiovascular disease, various cancers and nervous disease. NF-κB can modulate the expression activity of other transcriptional factors. Inhibition of IKK and IκBα phosphorylation, the decrease of nuclear translocation of p65 and the reduction of intracellular glutathione level determined the up-regulation or down-regulation of expression of NF-κB target genes.
    [Show full text]
  • SL10302R-FITC.Pdf
    SunLong Biotech Co.,LTD Tel: 0086-571- 56623320 Fax:0086-571- 56623318 E-mail:[email protected] www.sunlongbiotech.com Rabbit Anti-Hrk/FITC Conjugated antibody SL10302R-FITC Product Name: Anti-Hrk/FITC Chinese Name: FITC标记的BCL2相互作用蛋白质抗体 Activator of apoptosis harakiri; Activator of apoptosis Hrk; BCL2 interacting protein; BH3 interacting domain protein 3; BID3; DP5; Harakiri BCL2 interacting protein; Alias: Human brain 3UTR of mRNA for neuronal death protein partial sequence; Neuronal death protein DP5; HRK_HUMAN. Organism Species: Rabbit Clonality: Polyclonal React Species: Human,Mouse,Rat,Dog,Cow, ICC=1:50-200IF=1:50-200 Applications: not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. Molecular weight: 10kDa Form: Lyophilized or Liquid Concentration: 1mg/ml immunogen: KLH conjugated synthetic peptide derived from human Activator of apoptosis harakiri Lsotype: IgG Purification: affinitywww.sunlongbiotech.com purified by Protein A Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year Storage: when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. background: This gene encodes a member of the BCL-2 protein family. Members of this family are involved in activating or inhibiting apoptosis. The encoded protein localizes to Product Detail: intracellular membranes. This protein promotes apoptosis by interacting with the apoptotic inhibitors BCL-2 and BCL-X(L) via its BH3 domain.
    [Show full text]
  • In Vitro Targeting of Transcription Factors to Control the Cytokine Release Syndrome in 2 COVID-19 3
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.29.424728; this version posted December 30, 2020. 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 4.0 International license. 1 In vitro Targeting of Transcription Factors to Control the Cytokine Release Syndrome in 2 COVID-19 3 4 Clarissa S. Santoso1, Zhaorong Li2, Jaice T. Rottenberg1, Xing Liu1, Vivian X. Shen1, Juan I. 5 Fuxman Bass1,2 6 7 1Department of Biology, Boston University, Boston, MA 02215, USA; 2Bioinformatics Program, 8 Boston University, Boston, MA 02215, USA 9 10 Corresponding author: 11 Juan I. Fuxman Bass 12 Boston University 13 5 Cummington Mall 14 Boston, MA 02215 15 Email: [email protected] 16 Phone: 617-353-2448 17 18 Classification: Biological Sciences 19 20 Keywords: COVID-19, cytokine release syndrome, cytokine storm, drug repurposing, 21 transcriptional regulators 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.12.29.424728; this version posted December 30, 2020. 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 4.0 International license. 22 Abstract 23 Treatment of the cytokine release syndrome (CRS) has become an important part of rescuing 24 hospitalized COVID-19 patients. Here, we systematically explored the transcriptional regulators 25 of inflammatory cytokines involved in the COVID-19 CRS to identify candidate transcription 26 factors (TFs) for therapeutic targeting using approved drugs.
    [Show full text]
  • Genome-Wide Analysis of the Zebrafish ETS Family Identifies Three Genes Required for Hemangioblast Differentiation Or Angiogenesis
    Genome-Wide Analysis of the Zebrafish ETS Family Identifies Three Genes Required for Hemangioblast Differentiation or Angiogenesis Feng Liu, Roger Patient Abstract—ETS domain transcription factors have been linked to hematopoiesis, vasculogenesis, and angiogenesis. However, their biological functions and the mechanisms of action, remain incompletely understood. Here, we have performed a systematic analysis of zebrafish ETS domain genes and identified 31 in the genome. Detailed gene expression profiling revealed that 12 of them are expressed in blood and endothelial precursors during embryonic development. Combined with a phylogenetic tree assay, this suggests that some of the coexpressed genes may have redundant or additive functions in these cells. Loss-of-function analysis of 3 of them, erg, fli1, and etsrp, demonstrated that erg and fli1 act cooperatively and are required for angiogenesis possibly via direct regulation of an endothelial cell junction molecule, VE-cadherin, whereas etsrp is essential for primitive myeloid/endothelial progenitors (hemangio- blasts) in zebrafish. Taken together, these results provide a global view of the ETS genes in the zebrafish genome during embryogenesis and provide new insights on the functions and biology of erg, fli1, and etsrp, which could be applicable to higher vertebrates, including mice and humans. (Circ Res. 2008;103:1147-1154.) Key Words: zebrafish Ⅲ gene duplication Ⅲ ETS transcription factors Ⅲ hemangioblast Ⅲ angiogenesis ebrafish has been recognized as an excellent genetic and during embryonic development and adulthood and have been Zdevelopmental biology model to study hematopoiesis linked with diverse biological processes, from hematopoiesis, and vessel development. Large numbers of genetic mutants vasculogenesis, and angiogenesis to neurogenesis. Many and transgenic lines in both blood and endothelial lineages important blood and endothelial regulators have well- have become available.
    [Show full text]
  • Differential Gene Expression in Brain and Liver Tissue of Wistar Rats After
    Article Differential Gene Expression in Brain and Liver Tissue of Wistar Rats after Rapid Eye Movement Sleep Deprivation Atul Pandey 1,2,* , Ryan Oliver 2 and Santosh K Kar 1,3,* 1 School of Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India 2 Department of Ecology, Evolution, and Behavior, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel; [email protected] 3 Nano Herb Research Laboratory, Kalinga Institute of Industrial Technology (KIIT) Technology Bio Incubator, Campus-11, KIIT Deemed to be University, Bhubaneswar, Odisha 751024, India * Correspondence: [email protected] (A.P.); santoshkariis@rediffmail.com (S.K.K.); Tel.: +972-547301848 (A.P.); +91-9937085111 (S.K.K.) Received: 10 September 2020; Accepted: 21 October 2020; Published: 23 October 2020 Abstract: Sleep is essential for the survival of most living beings. Numerous researchers have identified a series of genes that are thought to regulate “sleep-state” or the “deprived state”. As sleep has a significant effect on physiology, we believe that lack of total sleep, or particularly rapid eye movement (REM) sleep, for a prolonged period would have a profound impact on various body tissues. Therefore, using the microarray method, we sought to determine which genes and processes are affected in the brain and liver of rats following nine days of REM sleep deprivation. Our findings showed that REM sleep deprivation affected a total of 652 genes in the brain and 426 genes in the liver. Only 23 genes were affected commonly, 10 oppositely, and 13 similarly across brain and liver tissue.
    [Show full text]
  • Analysis of Gene Expression in Wild Type and Notch1 Mutant Retinal Cells by Single Cell Profiling Karolina Mizeracka1, Jeffrey M
    Research Article Developmental Dynamics DOI 10.1002/dvdy.24006 Analysis of gene expression in wild type and Notch1 mutant retinal cells by single cell profiling Karolina Mizeracka1, Jeffrey M. Trimarchi1,2, Michael B. Stadler3, Constance L. Cepko1,4 1 Department of Genetics, Department of Ophthalmology, Harvard Medical School, Boston, MA 02115 2 Current Address: Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50014 3 Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland 4 Howard Hughes Medical Institute, Department of Genetics, Department of Ophthalmology, Harvard Medical School, Boston, MA 02115 Correspondence: Constance Cepko 77 Avenue Louis Pasteur, Boston, MA 02115 Phone: (617) 432-7618 Fax: (617) 432-7595 Running title: Single cell profiling of Notch1 mutant retinal cells Key words: retina, progenitor, microarray, cell fate Summary: • Profiling of individual Notch1 deficient and wild type postnatal retinal cells on microarrays reveals changes in gene expression obscured by whole tissue analysis • Notch1 deficient cells downregulate progenitor and cell cycle markers with a concomitant upregulation in early rod photoreceptor markers • Based on classification, single Notch1 deficient and wild type cells represent Developmental Dynamics transition from progenitor to postmitotic cell • Individual wild type retinal cells express cell type markers of both photoreceptors and interneurons Grant sponsor and number: National Institutes of Health Grant R01EY09676 Accepted Articles are accepted, unedited articles for future issues, temporarily published online in advance of the final edited version. © 2013 Wiley Periodicals, Inc. Received: Mar 04, 2013; Revised: May 02, 2013; Accepted: May 13, 2013 Developmental Dynamics Page 2 of 66 Abstract Background: The vertebrate retina comprises sensory neurons, the photoreceptors, as well as many other types of neurons and one type of glial cell.
    [Show full text]