DOCSLIB.ORG

A Strategy for Enhancing the Transcriptional Activity of Weak Cell Type-Specific Promoters

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A Strategy for Enhancing the Transcriptional Activity of Weak Cell Type-Specific Promoters Gene Therapy (1998) 5, 1656–1664 1998 Stockton Press All rights reserved 0969-7128/98 $12.00 http://www.stockton-press.co.uk/gt A strategy for enhancing the transcriptional activity of weak cell type-specific promoters DM Nettelbeck, V Je´roˆme and R Mu¨ ller Institut fu¨r Molekularbiologie und Tumorforschung (IMT), Philipps-Universita¨t Marburg, Emil-Mannkopff-Strasse 2, D-35033 Marburg, Germany Cell type- and tissue-specific promoters play an important moter to drive the simultaneous expression of the desired role in the development of site-selective vectors for gene effector/reporter gene product and a strong artificial tran- therapy. A large number of highly specific promoters has scriptional activator which stimulates transcription through been described, but their applicability is often hampered appropriate binding sites in the promoter. Using a VP16- by their inefficient transcriptional activity. In this study, we LexA chimeric transcription factor, we show that this describe a new strategy for enhancing the activity of weak approach leads to a 14- to Ͼ100-fold enhancement of both promoters without loss of specificity. The basic principle of the endothelial cell-specific von Willebrand factor promoter this strategy is to establish a positive feedback loop which and the gastrointestinal-specific sucrase-isomaltase pro- is initiated by transcription from a cell type-specific pro- moter while maintaining approximately 30- to Ͼ100-fold moter. This was achieved by using a cell type-specific pro- cell type specificity. Keywords: tissue-specific promoters; endothelial cell-specific transcription; von Willebrand factor promoter; gastrointes- tinal-specific transcription; sucrase isomaltase promoter; transcriptional targeting Introduction most, if not all, tumor cells have been successfully incor- porated into experimental gene therapy strategies. Fur- The construction of vectors targeting gene expression to thermore, regulatory sequences inducible by therapeutic specific sites in the organism is one of the current chal- dose of ionizing radiation, such as the egr-1 promoter,13 lenges in the field of gene therapy. In view of the fact or in response to oncogenic lesions, such as the erbB2 that the efficient targeted transduction of genes still rep- promoter,14 are worth mentioning in the same context. In resents a major obstacle, the possibility of restricting gene addition to these promoters, a plethora of other poten- expression to specific cell populations through specific tially useful transcriptional regulatory sequences with a 1–3 promoters is of particular importance. This not only high degree of specificity has been described.1–3 applies to the treatment of systemic diseases, such as Some of these promoters appear to be ideally suited metastatic cancer, but also to local gene therapy, whose for gene therapy, because they combine strong transcrip- efficacy and safety can be improved by the use of cell tional activity with a high degree of specificity. This is, type-specific promoters which keep the expression of the for instance, true for the melanocyte-specific tyrosinase therapeutic gene in non-target cells at a minimum. promoter.4–6,15 Frequently, however, specific promoters For these reasons, many of the recently described are inefficient activators of transcription which severely experimental gene therapy protocols make use of cell limits their applicability. A typical example is the von 1–3 type- or tissue-specific promoters. These include the Willebrand factor (vWF) promoter, which exhibits a tyrosinase promoter for directing gene expression to particularly high degree of endothelial cell (EC) speci- 4–6 ␣ melanoma cells, the -fetoprotein promoter for hepa- ficity compared with other EC promoters, but is a poor 7 toma cells, the mucin-1 promoter for mammary carci- activator of transcription16–19 (see also Results below). 8 noma, the prostate-specific antigen promoter for prostate Other EC promoters are stronger, but less specific, such 9 carcinoma or the neuron-specific enolase promoter for as the promoters for PECAM-1/CD3120,21 or flk- 10 neuronal cells. In other studies, transcriptional control 1/KDR.22,23 Another example is the promoter of the suc- elements which can stimulate transcription in response rase-isomaltase (SI) gene which is highly specific for to disease-specific alterations, such as hypoxic con- intestinal cells and gastrointestinal tumors,24,25 but again 11 12 ditions or the loss of cell cycle checkpoints in tumors, is a poor transcriptional activator (see Results below). In have also been used. In these scenarios, promoters con- the present study, we have chosen the vWF and SI pro- taining binding sites for hypoxia-inducible factor-1 (HIF- moters as models for the establishment of a strategy that 1) or the transcription factor E2F whose repression would be suitable for enhancing the transcriptional through the retinoblastoma protein pathway is lost in activity of weak promoters without affecting their speci- ficity (self-enhancing promoters).26 This goal was achi- eved by incorporating into these promoters a positive 26 Correspondence: R Mu¨ller feedback loop provided by a chimeric transcription fac- Received 20 May 1998; accepted 15 July 1998 tor consisting of the strong herpes simplex virus VP16 Enhancing cell type-specific transcription DM Nettelbeck et al 1657 transcriptional activation domain27,28 fused to the DNA- binding domain of LexA.29 Results Comparative analysis of EC-specific promoters A comparison of different EC promoters in various ECs and non-endothelial cell lines was performed in order to identify a suitable promoter for the transcriptional tar- geting of ECs. The promoters of the human vWF,17 PECAM-120,21 and KDR22 genes were analyzed in transi- ent luciferase assays using human umbilical vein ECs (HUVECs), bovine aortic ECs (BAECs), mouse NIH3T3 fibroblasts and the human melanoma cell line MeWo. As shown in Figure 1a, a clear selectivity for ECs was found with all three promoters, with the highest degree of cell type specificity seen with the vWF promoter. This is particularly well documented by the data in Figure 1b which show a complete lack of activity in all non-endo- thelial cell lines tested (NIH3T3, MeWo, 10T1/2, HeLa), ie RLU values similar to those obtained with the pro- moterless basic vector, while both the PECAM-1 and KDR promoter showed readily detectable activities in the non-endothelial cells (Figure 1a). We also tested other fragments of the KDR and PECAM-1 promoters (see Materials and methods for details), but these showed even lower levels of activity and/or specificity (data not shown). In contrast to its high degree of specificity, the vWF promoter is endowed with very poor transcriptional activation properties which severely limits its applica- bility (10–20% of SV40 promoter; Figure 1a). In a first attempt to improve the activity of the vWF promoter we tested a construct in which the SV40 enhancer was placed upstream of the promoter. This construct (SV40EvWF) showed strongly increased transcriptional activity, but the cell type specificity was largely lost. We also tested other EC promoters, including those of the human endo- glin (kindly provided by W. Graulich, IMT, Marburg, Figure 1 (a)Activity of different promoters in ECs and non-endothelial Germany) and human P-selectin30 genes, but these did cells after transient transfection of luciferase reporter constructs. Values not provide the same high degree of specificity seen with were standardized individually for each cell line using a SV40 promoter the vWF promoter (data not shown). We did not include (SV40p) construct. Basic, promoterless pGL3 vector; SV40EvWF, human the preproendothelin promoter31 in these studies because vWF promoter with an upstream SV40 enhancer. (b) Activity of the vWF promoter in ECs and non-endothelial cells after transient transfection of its activity has been reported to be restricted to bigger luciferase reporter constructs. Values were standardized individually for 32 blood vessels and is also active in some epithelial cells, each cell line using the promoterless pGL3 vector (basic) which allows for which precludes its use for targeting the vasculature in a clearer representation of the vWF promoter activity in the various cell tumors or other proliferative diseases. lines (especially the lack of activity in non-endothelial cells). The data shown are averages of triplicates. The error bars indicate the standard deviations. Establishment of a positive feedback loop for the vWF promoter: proof of principle In view of the data discussed above, we sought to estab- for details) into HUVECs, BAECs, NIH3T3 cells and lish a new strategy that would allow for an enhancement MeWo cells. The data in Figure 3b (standardized to SV40 of promoter activity by introducing a positive feedback promoter activity (SP40p) for each cell line) and Figure 3c loop. The basic principle of this strategy is to use a cell demonstrate that (1) the insertion of the seven LexA sites type-specific promoter to drive the simultaneous into the vWF promoter (LexvWFLuc) had no effect on expression of the desired effector/reporter gene product activity or specificity; (2) the cotransfection of a SV40 pro- and a strong artificial transcriptional activator, ie a chim- moter-driven expression vector encoding a VP16-LexA eric protein consisting of the DNA-binding domain of fusion protein (SV-VPLex) led to a dramatic increase in LexA29 and the strong herpes simplex virus VP16 tran- activity (approximately 30- to 70-fold), but the promoter scriptional activation
Load more

Recommended publications
  • Identification of the Promoter and a Transcriptional Enhancer of The
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 11356-11360, December 1993 Developmental Biology Identification of the promoter and a transcriptional enhancer of the gene encoding L-CAM, a calcium-dependent cell adhesion molecule (gene expression/regulatory sequences/morphogenesis/cadherins) BARBARA C. SORKIN, FREDERICK S. JONES, BRUCE A. CUNNINGHAM, AND GERALD M. EDELMAN The Department of Neurobiology, The Scripps Research Institute, 10666 North Torrey Pines Road, La Jolla, CA 92037 Contributed by Gerald M. Edelman, August 20, 1993 ABSTRACT L-CAM is a calcium-dependent cell adhesion expressed in the dermis, but rather in the adjacent epidermis molecule that is expressed in a characteristic place-dependent (6). pattern during development. Previous studies of ectopic ex- L-CAM mediates calcium-dependent cell-cell adhesion (7) pression of the chicken L-CAM gene under the control of via a homophilic mechanism; i.e., L-CAM on one cell binds heterologous promoters in transgenic mice suggested that directly to L-CAM on apposing cells (8). In all tissues where cis-acting sequences controlling the spatiotemporal expression the molecule is expressed, L-CAM is detected as a trans- patterns ofL-CAM were present within the gene itself. We have membrane protein of 124 kDa (7). Mammalian proteins now examined the L-CAM gene for sequences that control its uvomorulin (9), E-cadherin (10), cell-CAM 120/80 (11), and expression and have found an enhancer within the second Arcl (12) are similar to L-CAM in their biochemical and intron of the gene. A 2.5-kb Kpn I-EcoRI fragment from the functional properties and tissue distributions.
    [Show full text]
  • Molecular Basis of the Function of Transcriptional Enhancers
    cells Review Molecular Basis of the Function of Transcriptional Enhancers 1,2, 1, 1,3, Airat N. Ibragimov y, Oleg V. Bylino y and Yulii V. Shidlovskii * 1 Laboratory of Gene Expression Regulation in Development, Institute of Gene Biology, Russian Academy of Sciences, 34/5 Vavilov St., 119334 Moscow, Russia; [email protected] (A.N.I.); [email protected] (O.V.B.) 2 Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, 34/5 Vavilov St., 119334 Moscow, Russia 3 I.M. Sechenov First Moscow State Medical University, 8, bldg. 2 Trubetskaya St., 119048 Moscow, Russia * Correspondence: [email protected]; Tel.: +7-4991354096 These authors contributed equally to this study. y Received: 30 May 2020; Accepted: 3 July 2020; Published: 5 July 2020 Abstract: Transcriptional enhancers are major genomic elements that control gene activity in eukaryotes. Recent studies provided deeper insight into the temporal and spatial organization of transcription in the nucleus, the role of non-coding RNAs in the process, and the epigenetic control of gene expression. Thus, multiple molecular details of enhancer functioning were revealed. Here, we describe the recent data and models of molecular organization of enhancer-driven transcription. Keywords: enhancer; promoter; chromatin; transcriptional bursting; transcription factories; enhancer RNA; epigenetic marks 1. Introduction Gene transcription is precisely organized in time and space. The process requires the participation of hundreds of molecules, which form an extensive interaction network. Substantial progress was achieved recently in our understanding of the molecular processes that take place in the cell nucleus (e.g., see [1–9]).
    [Show full text]
  • Promoter Architecture and Sex-Specific Gene Expression In
    | INVESTIGATION Promoter Architecture and Sex-Specific Gene Expression in Daphnia pulex R. Taylor Raborn,*,†,1 Ken Spitze,* Volker P. Brendel,*,†,2 and Michael Lynch*,2 *Department of Biology and †School of Informatics and Computing, Indiana University, Bloomington, Indiana 47405 ORCID IDs: 0000-0001-6249-8033 (R.T.R.); 0000-0003-4289-6637 (K.S.); 0000-0002-8055-7508 (V.P.B.) ABSTRACT Large-scale transcription start site (TSS) profiling produces a high-resolution, quantitative picture of transcription initiation and core promoter locations within a genome. However, application of TSS profiling to date has largely been restricted to a small set of prominent model systems. We sought to characterize the cis-regulatory landscape of the water flea Daphnia pulex, an emerging model arthropod that reproduces both asexually (via parthenogenesis) and sexually (via meiosis). We performed Cap Analysis of Gene Expression (CAGE) with RNA isolated from D. pulex within three developmental states: sexual females, asexual females, and males. Identified TSSs were utilized to generate a “Daphnia Promoter Atlas,” i.e., a catalog of active promoters across the surveyed states. Analysis of the distribution of promoters revealed evidence for widespread alternative promoter usage in D. pulex, in addition to a prominent fraction of compactly-arranged promoters in divergent orientations. We carried out de novo motif discovery using CAGE- defined TSSs and identified eight candidate core promoter motifs; this collection includes canonical promoter elements (e.g., TATA and Initiator) in addition to others lacking obvious orthologs. A comparison of promoter activities found evidence for considerable state- specific differential gene expression between states. Our work represents the first global definition of transcription initiation and promoter architecture in crustaceans.
    [Show full text]
  • Escherichia Coli (Gene Fusion/Attenuator/Terminator/RNA Polymerase/Ribosomal Proteins) GERARD BARRY, CATHERINE L
    Proc. Natl. Acad. Sci. USA Vol. 76, No. 10, pp. 4922-4926, October 1979 Biochemistry Control features within the rplJL-rpoBC transcription unit of Escherichia coli (gene fusion/attenuator/terminator/RNA polymerase/ribosomal proteins) GERARD BARRY, CATHERINE L. SQUIRES, AND CRAIG SQUIRES Department of Biological Sciences, Columbia University, New York, New York 10027 Communicated by Cyrus Levinthal, July 2, 1979 ABSTRACT Gene fusions constructed in vitro have been regulation could occur (4-7). Yet under certain conditions, used to examine transcription regulatory signals from the operon coordinate of the RNA which encodes ribosomal proteins L10 and L7/12 and the RNA expression polymerase subunits and polymerase P and #I subunits (the rplJL-rpoBC operon). Por- ribosomal proteins is not observed. This is especially true of the tions of this operon, which were obtained by in vitro deletions, rplJL-rpoBC transcription unit which encodes the ribosomal have been placed between the ara promoter and the lacZgene proteins L10 and L7/12 and the RNA polymerase subunits 13 in the gene-fusion plasmid pMC81 developed by M. Casadaban and 13'. For example, only the ribosomal proteins are modulated and S. Cohen. The effect of the inserted DNA segment on the by the stringent regulation system (8, 9) whereas a transient expression of the IacZ gene (in the presence and absence of arabinose) permits the localization of regulatory signals to dis- stimulatory effect of rifampicin is specific for the RNA poly- crete regions of the rpIJL-rpoBC operon. An element that re- merase subunits (10, 11). In addition, different amounts of duces the level of distal gene expression to one-sixth is located mRNA hybridize to the rpl and rpo regions of the rplJL-rpoBC on a fragment which spans the rplL-rpoB intercistronic region.
    [Show full text]
  • Promoter RNA Sequencing
    www.nature.com/scientificreports OPEN Promoter RNA sequencing (PRSeq) for the massive and quantitative promoter analysis in vitro Received: 7 August 2018 Shoji Ohuchi1,3, Thorsten Mascher 1 & Beatrix Suess2 Accepted: 1 February 2019 Analysis of promoter strength and specifcity is important for understanding and engineering gene Published: xx xx xxxx regulation. Here, we report an in vitro promoter analysis method that can achieve both massiveness and quantitativeness. In this approach, a pool of single-stranded DNA with a partially randomized promoter sequence to be analyzed is chemically synthesized. Through enzymatic reactions, the randomized sequence will be copied to the downstream region, resulting in a template DNA pool that carries its own promoter information on its transcribed region. After in vitro transcription of the DNA pool with an RNA polymerase of interest, the sequences of the resulting transcripts will be analyzed. Since the promoter strength linearly correlates to the copy number of transcript, the strength of each promoter sequence can be evaluated. A model experiment of T7 promoter variants demonstrated the quantitativeness of the method, and the method was applied for the analysis of the promoter of cyanophage Syn5 RNA polymerase. This method provides a powerful approach for analyzing the complexity of promoter specifcity and discrimination for highly abundant and often redundant alternative sigma factors such as the extracellular function (ECF) sigma factors. Transcription initiation is the key step for controlling gene expression especially in bacterial and archaeal cells. Tus, analysis of promoter strength and specifcity is important for understanding gene regulation. Traditionally, promoter analysis is performed employing in vivo reporter gene fusions [reviewed in1].
    [Show full text]
  • CG-3'-Rich Region in the Promoter of the Transcriptionally Frequently Silenced RET Protooncogene Lacks Methylated Cytidine Residues
    Oncogene (1998) 17, 2573 ± 2583 ã 1998 Stockton Press All rights reserved 0950 ± 9232/98 $12.00 http://www.stockton-press.co.uk/onc A5'-CG-3'-rich region in the promoter of the transcriptionally frequently silenced RET protooncogene lacks methylated cytidine residues Marc Munnes1, Giovanna Patrone2, Birgit Schmitz1, Giovanni Romeo2 and Walter Doer¯er1 1Institut fuÈr Genetik, UniversitaÈtzuKoÈln, D-50931 KoÈln, Germany; and 2UniversitaÁ di Genova, FacoltaÁ di Medicina e Laboratorio di Genetica Molecolare, Istituto G. Gaslini, I-16148 Genova, Italy In a large proportion of familial and sporadic cases of Keywords: HSCR patients; RET protooncogene Hirschsprung disease (HSCR) mutations in the RET promoter; 5'-CG-3'-rich region in the RET promoter (rearranged during transfection) protooncogene have been described. We have investigated the structure of the RET gene promoter and have analysed a region of approximately 1000 nucleotides in its promoter and 5'- Introduction upstream segments for the occurrence of 5-methyldeoxy- cytidine (5-mC) residues by using the bisul®te protocol of The human RET protooncogene is controlled by a the genomic sequencing method. With an estimated promoter harboring several transcription factor sensitivity of about 93% of this technique, not a single binding sites (Itoh et al., 1992), such as four 5-mC residue could be detected in the control region of a tandemly repeated GC-boxes (Dynan and Tjian, gene that seems to be silenced or exhibit low activity in 1985), an ETF binding site (Kageyama et al., 1989) many adult tissues. In these experiments, the DNAs of and Sp1 and AP-2 binding sites. Most of these sites peripheral white blood cells (PWBC) from four healthy are located within the 5'-CpG-3'-rich region in individuals, from seven patients with familial HSCR, as proximity to the transcriptional start site.
    [Show full text]
  • Regulation of RNA Polymerase II Transcription
    Regulation of RNA polymerase II transcription Ronny Drapkin, Alejandro Merino and Danny Reinberg Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, USA Transcription initiation plays a central role in the regulation of gene expression. Exciting developments in the last year have furthered our understanding of the interactions between general transcription factors and how these factors respond to modulators of transcription. Current Opinion in Cell Biology 1993, 5:469-476 Introduction TFIIJ. Formation of the DAB--polFEHJ complex, in the presence of each of four ribonucleoside triphosphates, Cellular growth and differentiation employ precise mech- enables RNAPII to clear the promoter region and initiate anisms to regulate the expression of various genes. One RNA synthesis from a specific start site [ 51. of the most rudimentary mechanisms for a cell to control The past year has seen intense activity aimed at elucidat- the functional levels of a protein is to modulate the lev- ing the molecular mechanisms underlying transcription els of mRNA encoding that polypeptide. It is therefore not initiation. In particular, the interactions that GTFs can surprising that most of the genetic programs that main- mediate, the GTF requirements for initiation, the role tain the cell in a constant state of flux mediate their effects of RNAPII phosphorylation, and the phenomenon of by impinging on mechanisms that control transcription antirepression in the process of activation have been initiation. the subject of many studies. These most recent develop- In contrast to prokaryotic RNA polymerase, eukaryotic ments are the focus of this review. enzymes require multiple accessory proteins to acquire promoter specificity.
    [Show full text]
  • Saccharomyces Cerevisiae Promoter Engineering Before and During the Synthetic Biology Era
    biology Review Saccharomyces cerevisiae Promoter Engineering before and during the Synthetic Biology Era Xiaofan Feng and Mario Andrea Marchisio * School of Pharmaceutical Science and Technology, Tianjin University, 92 Weijin Road, Tianjin 300072, China; [email protected] * Correspondence: [email protected] or [email protected] Simple Summary: Promoters are DNA sequences where the process of transcription starts. They can work constitutively or be controlled by environmental signals of different types. The quantity of proteins and RNA present in yeast genetic circuits highly depends on promoter strength. Hence, they have been deeply studied and modified over, at least, the last forty years, especially since the year 2000 when Synthetic Biology was born. Here, we present how promoter engineering changed over these four decades and discuss its possible future directions due to novel computational methods and technology. Abstract: Synthetic gene circuits are made of DNA sequences, referred to as transcription units, that communicate by exchanging proteins or RNA molecules. Proteins are, mostly, transcription factors that bind promoter sequences to modulate the expression of other molecules. Promoters are, therefore, key components in genetic circuits. In this review, we focus our attention on the construction of artificial promoters for the yeast S. cerevisiae, a popular chassis for gene circuits. We describe the initial techniques and achievements in promoter engineering that predated the start of the Synthetic Biology epoch of about 20 years. We present the main applications of synthetic Citation: Feng, X.; Marchisio, M.A. promoters built via different methods and discuss the latest innovations in the wet-lab engineering Saccharomyces cerevisiae Promoter of novel promoter sequences.
    [Show full text]
  • Biol. Pharm. Bull. 43(4): 742-746 (2020)
    742 Biol. Pharm. Bull. 43, 742–746 (2020) Vol. 43, No. 4 Note PRC2 Components Maintain DNA Hypermethylation of the Upstream Promoter and Regulate Robo4 Expression in Endothelial Cells Kohei Izawa,# Keisuke Shirakura,# Koji Kakiuchi, Nobuaki Funahashi, Naoki Maekawa, Nobumasa Hino, Toru Tanaka, Takefumi Doi, and Yoshiaki Okada* Graduate School of Pharmaceutical Sciences, Osaka University; 1–6 Yamadaoka, Suita, Osaka 565–0871, Japan. Received November 15, 2019; accepted January 24, 2020 Roundabout4 (Robo4) is an endothelial cell-specific protein that stabilizes the vasculature in pathologi- cal angiogenesis and inflammation. We previously determined a 3-kb Robo4 promoter and demonstrated the importance of the upstream region for nuclear factor-kappaB (NF-κB)-mediated promoter activation in- duced by tumor necrosis factor α (TNFα). This region contains unique genomic features, including promoter region-specific DNA hypermethylation and chromatin condensation; however, the function of the region re- mains poorly understood. In this study, we analyzed the DNA sequences of the region and identified a motif for polycomb repressive complex 2 (PRC2). Chromatin immunoprecipitation assay indicates the binding of the PRC2 component, SUZ12, to the motif. A mutation in the motif decreased DNA methylation in embryonic stem cells and increased Robo4 promoter activity in endothelial cells. An inhibitor for the PRC2 component, EZH2, induced the promoter activity and expression of Robo4 in endothelial cells treated with or without TNFα. Taken together, these
    [Show full text]
  • A Coreceptor for HIV-1 Entry CXCR4, Suppresses, the Promoter Activity Of
    USF/c-Myc Enhances, While Yin-Yang 1 Suppresses, the Promoter Activity of CXCR4, a Coreceptor for HIV-1 Entry This information is current as Masako Moriuchi, Hiroyuki Moriuchi, David M. Margolis of September 23, 2021. and Anthony S. Fauci J Immunol 1999; 162:5986-5992; ; http://www.jimmunol.org/content/162/10/5986 Downloaded from References This article cites 41 articles, 26 of which you can access for free at: http://www.jimmunol.org/content/162/10/5986.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 23, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 1999 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. USF/c-Myc Enhances, While Yin-Yang 1 Suppresses, the Promoter Activity of CXCR4, a Coreceptor for HIV-1 Entry Masako Moriuchi,1* Hiroyuki Moriuchi,1,2* David M. Margolis,† and Anthony S. Fauci* Transcription factors USF1 and USF2 up-regulate gene expression (i.e., HIV-1 long terminal repeats) via interaction with an E box on their target promoters, which is also a binding site for c-Myc.
    [Show full text]
  • EZH2 Regulates Neuroblastoma Cell Differentiation Via NTRK1 Promoter Epigenetic Modifications
    Oncogene (2018) 37:2714–2727 https://doi.org/10.1038/s41388-018-0133-3 ARTICLE EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications 1,2 1 3,4 3,5 1 Zhenghao Li ● Hisanori Takenobu ● Amallia Nuggetsiana Setyawati ● Nobuhiro Akita ● Masayuki Haruta ● 1 1 1,6 1 1 1 Shunpei Satoh ● Yoshitaka Shinno ● Koji Chikaraishi ● Kyosuke Mukae ● Jesmin Akter ● Ryuichi P. Sugino ● 7 8 9 1 1,2 Atsuko Nakazawa ● Akira Nakagawara ● Hiroyuki Aburatani ● Miki Ohira ● Takehiko Kamijo Received: 25 May 2017 / Revised: 20 October 2017 / Accepted: 27 November 2017 / Published online: 6 March 2018 © The Author(s) 2018. This article is published with open access Abstract The polycomb repressor complex 2 molecule EZH2 is now known to play a role in essential cellular processes, namely, cell fate decisions, cell cycle regulation, senescence, cell differentiation, and cancer development/progression. EZH2 inhibitors have recently been developed; however, their effectiveness and underlying molecular mechanisms in many malignancies have not yet been elucidated in detail. Although the functional role of EZH2 in tumorigenesis in neuroblastoma (NB) has been investigated, mutations of EZH2 have not been reported. A Kaplan–Meier analysis on the event free survival and 1234567890();,: overall survival of NB patients indicated that the high expression of EZH2 correlated with an unfavorable prognosis. In order to elucidate the functional roles of EZH2 in NB tumorigenesis and its aggressiveness, we knocked down EZH2 in NB cell lines using lentivirus systems. The knockdown of EZH2 significantly induced NB cell differentiation, e.g., neurite extension, and the neuronal differentiation markers, NF68 and GAP43.
    [Show full text]
  • Cis-Acting Elements Responsible for Muscle-Specific Expression of the Myosin Heavy Chain ,3 Gene
    kQ-D 1992 Oxford University Press Nucleic Acids Research, Vol. 20, No. 7 1793-1799 Cis-acting elements responsible for muscle-specific expression of the myosin heavy chain ,3 gene Noriko Shimizul +, Gwen Prior4, Patrick K.Umeda5 and Radovan Zak' 2S* Departments of 1Medicine, 2Organismal Biology and Anatomy, and 3Pharmacological & Physiological Sciences, The University of Chicago, Chicago, IL 60637, 4Marine Biology Laboratory, Woods Hole, MA 02543 and 5Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA Received August 8, 1991; Revised and Accepted February 19, 1992 ABSTRACT The 5' flanking region of the rabbit myosin heavy chain (1). In mammals, one of the members, the myosin HC , gene, (HC) a gene extending 295 bp upstream from the cap is expressed early during development in skeletal muscles and site provides muscle-specific transcriptional activity. In becomes restricted to slow twitch fibers in adults. The myosin this study, we have identified and functionally HC is also a major myosin HC isoform in cardiac muscle. The characterized cis-acting elements that regulate the differential expression of the HC ,3 gene, like that of most of muscle-specific expression within this region. By using the other muscle-specific genes, is regulated primarily by linker-scanner (LS) mutants between - 295 bp and a transcriptional mechanisms (2), indicating that the interaction of putative TATA box, we found five distinct positive cis- the cis-acting regulatory elements with the trans-acting nuclear acting sequences necessary for transcription: element factors plays an important role in determining the specific pattern A, the sequences between -276 and -263, which of transcriptional regulation.
    [Show full text]