DOCSLIB.ORG

MSX1 Modulates RLR-Mediated Innate Antiviral Signaling by Facilitating Assembly of TBK1-Associated Complexes

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
MSX1 Modulates RLR-Mediated Innate Antiviral Signaling by Facilitating Assembly of TBK1-Associated Complexes MSX1 Modulates RLR-Mediated Innate Antiviral Signaling by Facilitating Assembly of TBK1-Associated Complexes This information is current as Liu-Ting Chen, Ming-Ming Hu, Zhi-Sheng Xu, Yu Liu and of October 1, 2021. Hong-Bing Shu J Immunol published online 18 May 2016 http://www.jimmunol.org/content/early/2016/05/17/jimmun ol.1600039 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2016/05/18/jimmunol.160003 Material 9.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • 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 October 1, 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 © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published May 18, 2016, doi:10.4049/jimmunol.1600039 The Journal of Immunology MSX1 Modulates RLR-Mediated Innate Antiviral Signaling by Facilitating Assembly of TBK1-Associated Complexes Liu-Ting Chen, Ming-Ming Hu, Zhi-Sheng Xu, Yu Liu, and Hong-Bing Shu Recognition of viral dsRNA by the retinoic acid–inducible gene-1–like receptors (RLRs) triggers signaling cascades that lead to activation of the TBK1 kinase and transcription factor IFN regulatory factor 3, induction of downstream antiviral genes, and innate antiviral responses. In this study, we identified muscle segment homeobox1 (MSX1) as an important modulator of RLR- mediated signaling pathways. Knockdown or knockout of MSX1 significantly impaired Sendai virus–triggered activation of TBK1 and IFN regulatory factor 3, induction of downstream antiviral genes, and cellular antiviral responses. Interestingly, MSX1 was translocated from the nucleus to cytoplasm, particularly mitochondria upon infection of Sendai virus. Biochemcially, MSX1 was important for assembly of TBK1/IKK-related kinase-associated protein 1/TNFR-associated factor-associated NF-kB activator complexes. Our results suggest that MSX1 is an important component of RLR-mediated signaling and reveal mechanisms on innate immune responses against RNA viruses. The Journal of Immunology, 2016, 197: 000–000. Downloaded from he innate immune response against virus begins with rec- TBK1 was first identified as an IkB kinase that mediates IKK and ognition of viral nucleic acids by specific pattern-recognition NF-kB activation in response to growth factors (10–12). It was later T receptors. Cytosolic dsRNA from viral genome or its rep- identified as a virus-activated kinase that coordinates with IKKε to lication intermediates are mainly recognized by retinoic acid– phosphorylate transcription factors IRF3 and IRF7, leading to in- inducible gene-1 (RIG-I) and melanoma differentiation-associated duction of type I IFNs (13–17). With the exception of plasmacytoid http://www.jimmunol.org/ gene 5 (MDA5). Both RIG-I and MDA5 contain two N-terminal dendritic cells, most cells deficient for both TBK1 and IKKε fail to tandem caspase activation and recruitment domains and a produce type I IFNs in response to viral infection (14, 17). C-terminal DexD/H box RNA helicase domain, and thus they are Due to the vital role of TBK1 in virus-induced signaling, the collectively called RIG-I–like receptors (RLRs). The binding of molecular events underlying TBK1 activation have been heavily RLR with viral dsRNA induces conformational changes and olig- investigated. Multiple molecules have been reported to regulate omerization of RLR that activate virus-triggered signaling adaptor activation of TBK1 to ensure suitable immune responses against (VISA, also known as MAVS, IPS-1, and Cardif) on the mito- invaded viruses. Suppressor of IKKε functions as a physiological chondrial and peroxisomal membranes. VISA forms a signalsome suppressor of TBK1, as well as IKKε, by sequestering IKKε/TBK1 by recruiting and activating multiple signaling components, in- in inactive complexes (18). Nod-like receptor family pyrin domain- by guest on October 1, 2021 cluding WDR5, MITA (also known as STING), TAK1, TBK1, containing 4 promotes degradation of TBK1 by recruiting the E3 and/or IKKε kinases, leading to activation of transcription factors ligase d 4 (DTX4) to TBK1 and promoting K48-linked poly- IFN regulatory factor (IRF)3 and NF-kB and induction of type I ubiquitination of TBK1 (19). Dual-specificity tyrosine phosphory- IFNs (IFN-a and IFN-b) and inflammatory cytokines (1–9). Type lation–regulated kinase 2 phosphorylates TBK1 at Ser527,leadingto I IFNs further activate the JAK–STAT signal transduction path- recruitment of Nod-like receptor family pyrin domain-containing 4 ways, leading to transcriptional induction of a wide range of down- and DTX4 to degrade TBK1 (20). Glycogen synthase kinase 3b stream antiviral genes. potentiates activation of TBK1 by promoting TBK1 self-association and self-phosphorylation at S172 (21). Several adaptor molecules are also reported to modulate activation of TBK1, including TNFR- College of Life Sciences, Medical Research Institute, Collaborative Innovation Cen- ter for Viral Immunology, Wuhan University, Wuhan 430072, China associated factor-associated NF-kB activator (TANK) and IKK- related kinase-associated protein 1 (NAP1). Deficiency of TANK ORCIDs: 0000-0001-9962-5286 (L.-T.C.); 0000-0001-9667-7662 (M.-M.H.); 0000- 0001-7034-0666 (Z.-S.X.). or NAP1 inhibits Sendai virus (SeV)–induced activation of IRF3, Received for publication January 7, 2016. Accepted for publication April 22, 2016. suggesting that these adaptor proteins are required for RLR- This work was supported by Ministry of Science and Technology of China Grants mediated signaling. NAP1 connects RLRs with TBK1 to activate 2014CB910103 and 2012CB910201 and National Natural Science Foundation of IRF3 (22). Consistently, TANK forms complexes with VISA, China Grants 31521091, 91429304, and 31370867. TRAF3, and TBK1 (23). Therefore, NAP1 and TANK are adaptors Address correspondence and reprint requests to Dr. Hong-Bing Shu, College of Life linking TBK1 with the upstream signaling complexes, leading to Sciences, Medical Research Institute, Collaborative Innovation Center for Viral Immu- nology, Wuhan University, Wuhan 430072, China. E-mail address: [email protected] phosphorylation and activation of TBK1 (22, 23). The online version of this article contains supplemental material. Muscle segment homeobox1 (MSX1) is a member of the MSX transcription factors that play key roles in development by re- Abbreviations used in this article: HA, hemagglutinin; IRF, IFN regulatory factor; ISRE, IFN-stimulated regulatory element; MDA5, melanoma differentiation- pressing gene expression through interacting with components of associated gene 5; MSX1, muscle segment homeobox1; NAP1, IKK-related the core transcription complexes as well as other homeoproteins kinase-associated protein 1; NDV, Newcastle disease virus; poly (I:C), polyinosinic- polycytidylic acid; qPCR, quantitative PCR; RIG-I, retinoic acid–inducible gene-1; (24, 25). MSX1 plays important roles in multiple signaling path- RLR, RIG-I–like receptor; SeV, Sendai virus; TANK, TNFR-associated factor- ways, including Wnt/b-catenin and D-Notch pathways (26–28). associated NF-kB activator; TRAF, TNFR-associated factor; VISA, virus-triggered Mutations of MSX1 in both humans and mice have been associ- signaling adaptor; VSV, vesicular stomatitis virus. ated with impaired development of cranial neural crest-derived Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 structures, oral clefts, and nonsyndromic oligodontia (24, 29–31). www.jimmunol.org/cgi/doi/10.4049/jimmunol.1600039 2 MSX1 REGULATES RLR SIGNALING However, the roles of MSX1 in RLR-mediated signaling path- HCT116 were transfected by lipofectamine 2000 (Invitrogen). Empty ways have not been previously studied. control plasmid was added to ensure that each transfection received the In this study, we identified MSX1 as an important regulator of same amount of total DNA. To normalize for transfection efficiency, pRL- TK (Renilla luciferase) reporter plasmid (0.02 mg) was added to each RLR-mediated signaling pathways. Knockdown or knockout of transfection. Luciferase assays were performed using a dual-specific lu- MSX1 significantly inhibited RLR-mediated activation of IRF3, ciferase assay kit (Promega, Madison, WI). Firefly luciferase activities induction of downstream antiviral genes, and cellular antiviral were normalized on the basis of Renilla luciferase activities. responses. MSX1 was translocated from the nucleus to the cyto- Quantitative real-time PCR plasm upon viral infection and important for assembly of TBK1- associated complexes. Our findings reveal a previously unidentified Total RNA was isolated from cells using TRIzol reagent (Invitrogen). After reverse transcription with an oligo deoxy-thymine primer using a RevertAid function of MSX1 and provide insight into the mechanisms of First Strand cDNA Synthesis Kit (Fermentas), aliquots of products were innate antiviral responses. subjected to
Load more

Recommended publications
  • Ncomms2254.Pdf
    ARTICLE Received 3 May 2012 | Accepted 5 Nov 2012 | Published 4 Dec 2012 DOI: 10.1038/ncomms2254 The RB family is required for the self-renewal and survival of human embryonic stem cells Jamie F. Conklin1,2,3, Julie Baker2,3 & Julien Sage1,2,3 The mechanisms ensuring the long-term self-renewal of human embryonic stem cells are still only partly understood, limiting their use in cellular therapies. Here we found that increased activity of the RB cell cycle inhibitor in human embryonic stem cells induces cell cycle arrest, differentiation and cell death. Conversely, inactivation of the entire RB family (RB, p107 and p130) in human embryonic stem cells triggers G2/M arrest and cell death through functional activation of the p53 pathway and the cell cycle inhibitor p21. Differences in E2F target gene activation upon loss of RB family function between human embryonic stem cells, mouse embryonic stem cells and human fibroblasts underscore key differences in the cell cycle regulatory networks of human embryonic stem cells. Finally, loss of RB family function pro- motes genomic instability in both human and mouse embryonic stem cells, uncoupling cell cycle defects from chromosomal instability. These experiments indicate that a homeostatic level of RB activity is essential for the self-renewal and the survival of human embryonic stem cells. 1 Department of Pediatrics, Stanford Medical School, Stanford, California 94305, USA. 2 Department of Genetics, Stanford Medical School, Stanford, California 94305, USA. 3 Institute for Stem Cell Biology and Regenerative Medicine, Stanford Medical School, Stanford, California 94305, USA. Correspondence and requests for materials should be addressed to J.S.
    [Show full text]
  • Prospective Isolation of NKX2-1–Expressing Human Lung Progenitors Derived from Pluripotent Stem Cells
    The Journal of Clinical Investigation RESEARCH ARTICLE Prospective isolation of NKX2-1–expressing human lung progenitors derived from pluripotent stem cells Finn Hawkins,1,2 Philipp Kramer,3 Anjali Jacob,1,2 Ian Driver,4 Dylan C. Thomas,1 Katherine B. McCauley,1,2 Nicholas Skvir,1 Ana M. Crane,3 Anita A. Kurmann,1,5 Anthony N. Hollenberg,5 Sinead Nguyen,1 Brandon G. Wong,6 Ahmad S. Khalil,6,7 Sarah X.L. Huang,3,8 Susan Guttentag,9 Jason R. Rock,4 John M. Shannon,10 Brian R. Davis,3 and Darrell N. Kotton1,2 2 1Center for Regenerative Medicine, and The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA. 3Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, Texas, USA. 4Department of Anatomy, UCSF, San Francisco, California, USA. 5Division of Endocrinology, Diabetes and Metabolism, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA. 6Department of Biomedical Engineering and Biological Design Center, Boston University, Boston, Massachusetts, USA. 7Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. 8Columbia Center for Translational Immunology & Columbia Center for Human Development, Columbia University Medical Center, New York, New York, USA. 9Department of Pediatrics, Monroe Carell Jr. Children’s Hospital, Vanderbilt University, Nashville, Tennessee, USA. 10Division of Pulmonary Biology, Cincinnati Children’s Hospital, Cincinnati, Ohio, USA. It has been postulated that during human fetal development, all cells of the lung epithelium derive from embryonic, endodermal, NK2 homeobox 1–expressing (NKX2-1+) precursor cells.
    [Show full text]
  • Table S1 the Four Gene Sets Derived from Gene Expression Profiles of Escs and Differentiated Cells
    Table S1 The four gene sets derived from gene expression profiles of ESCs and differentiated cells Uniform High Uniform Low ES Up ES Down EntrezID GeneSymbol EntrezID GeneSymbol EntrezID GeneSymbol EntrezID GeneSymbol 269261 Rpl12 11354 Abpa 68239 Krt42 15132 Hbb-bh1 67891 Rpl4 11537 Cfd 26380 Esrrb 15126 Hba-x 55949 Eef1b2 11698 Ambn 73703 Dppa2 15111 Hand2 18148 Npm1 11730 Ang3 67374 Jam2 65255 Asb4 67427 Rps20 11731 Ang2 22702 Zfp42 17292 Mesp1 15481 Hspa8 11807 Apoa2 58865 Tdh 19737 Rgs5 100041686 LOC100041686 11814 Apoc3 26388 Ifi202b 225518 Prdm6 11983 Atpif1 11945 Atp4b 11614 Nr0b1 20378 Frzb 19241 Tmsb4x 12007 Azgp1 76815 Calcoco2 12767 Cxcr4 20116 Rps8 12044 Bcl2a1a 219132 D14Ertd668e 103889 Hoxb2 20103 Rps5 12047 Bcl2a1d 381411 Gm1967 17701 Msx1 14694 Gnb2l1 12049 Bcl2l10 20899 Stra8 23796 Aplnr 19941 Rpl26 12096 Bglap1 78625 1700061G19Rik 12627 Cfc1 12070 Ngfrap1 12097 Bglap2 21816 Tgm1 12622 Cer1 19989 Rpl7 12267 C3ar1 67405 Nts 21385 Tbx2 19896 Rpl10a 12279 C9 435337 EG435337 56720 Tdo2 20044 Rps14 12391 Cav3 545913 Zscan4d 16869 Lhx1 19175 Psmb6 12409 Cbr2 244448 Triml1 22253 Unc5c 22627 Ywhae 12477 Ctla4 69134 2200001I15Rik 14174 Fgf3 19951 Rpl32 12523 Cd84 66065 Hsd17b14 16542 Kdr 66152 1110020P15Rik 12524 Cd86 81879 Tcfcp2l1 15122 Hba-a1 66489 Rpl35 12640 Cga 17907 Mylpf 15414 Hoxb6 15519 Hsp90aa1 12642 Ch25h 26424 Nr5a2 210530 Leprel1 66483 Rpl36al 12655 Chi3l3 83560 Tex14 12338 Capn6 27370 Rps26 12796 Camp 17450 Morc1 20671 Sox17 66576 Uqcrh 12869 Cox8b 79455 Pdcl2 20613 Snai1 22154 Tubb5 12959 Cryba4 231821 Centa1 17897
    [Show full text]
  • Expression of the Human PAC1 Receptor Leads to Dose- Dependent Hydrocephalus-Related Abnormalities in Mice
    Expression of the human PAC1 receptor leads to dose- dependent hydrocephalus-related abnormalities in mice Bing Lang, … , Anthony J. Harmar, Sanbing Shen J Clin Invest. 2006;116(7):1924-1934. https://doi.org/10.1172/JCI27597. Research Article Neuroscience Hydrocephalus is a common and potentially devastating birth defect affecting the CNS, and its relationship with G protein–coupled receptors (GPCRs) is unknown. We have expressed 2, 4, or 6 copies of a GPCR — the human PAC1 receptor with a 130-kb transgene in the mouse nervous system in a pattern closely resembling that of the endogenous gene. Consistent with PAC1 actions, PKA and PKC activity were elevated in the brains of Tg mice. Remarkably, Tg mice developed dose-dependent hydrocephalus-like characteristics, including enlarged third and lateral ventricles and reduced cerebral cortex, corpus callosum, and subcommissural organ (SCO). Neuronal proliferation and apoptosis were implicated in hydrocephalus, and we observed significantly reduced neuronal proliferation and massively increased neuronal apoptosis in the developing cortex and SCO of Tg embryos, while neurite outgrowth and neuronal migration in vitro remain uncompromised. Ventricular ependymal cilia are crucial for directing cerebrospinal fluid flow, and ependyma of Tg mice exhibited disrupted cilia with increased phospho-CREB immunoreactivity. These data demonstrate that altered neuronal proliferation/apoptosis and disrupted ependymal cilia are the main factors contributing to hydrocephalus in PAC1-overexpressing mice. This is the first report to our knowledge demonstrating that misregulation of GPCRs can be involved in hydrocephalus-related neurodevelopmental disorders. Find the latest version: https://jci.me/27597/pdf Research article Related Commentary, page 1828 Expression of the human PAC1 receptor leads to dose-dependent hydrocephalus- related abnormalities in mice Bing Lang,1 Bing Song,1 Wendy Davidson,1 Alastair MacKenzie,1 Norman Smith,2 Colin D.
    [Show full text]
  • To Proliferate Or to Die: Role of Id3 in Cell Cycle Progression and Survival of Neural Crest Progenitors
    Downloaded from genesdev.cshlp.org on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press To proliferate or to die: role of Id3 in cell cycle progression and survival of neural crest progenitors Yun Kee and Marianne Bronner-Fraser1 Division of Biology, California Institute of Technology, Pasadena, California 91125, USA The neural crest is a unique population of mitotically active, multipotent progenitors that arise at the vertebrate neural plate border. Here, we show that the helix–loop–helix transcriptional regulator Id3 has a novel role in cell cycle progression and survival of neural crest progenitors in Xenopus. Id3 is localized at the neural plate border during gastrulation and neurulation, overlapping the domain of neural crest induction. Morpholino oligonucleotide-mediated depletion of Id3 results in the absence of neural crest precursors and a resultant loss of neural crest derivatives. This appears to be mediated by cell cycle inhibition followed by cell death of the neural crest progenitor pool, rather than a cell fate switch. Conversely, overexpression of Id3 increases cell proliferation and results in expansion of the neural crest domain. Our data suggest that Id3 functions by a novel mechanism, independent of cell fate determination, to mediate the decision of neural crest precursors to proliferate or die. [Keywords: Xenopus; Id3; neural crest; cell cycle; survival] Received September 1, 2004; revised version accepted January 19, 2005. The neural crest is an embryonic cell population that origi- et al. 2003), c-Myc (Bellmeyer et al. 2003), and Msx1 nates from the lateral edges of the neural plate during (Tribulo et al. 2003) have been identified in Xenopus nervous system formation.
    [Show full text]
  • Lncmyod Promotes Skeletal Myogenesis and Regulates Skeletal Muscle Fiber-Type Composition by Sponging Mir-370-3P
    G C A T T A C G G C A T genes Article LncMyoD Promotes Skeletal Myogenesis and Regulates Skeletal Muscle Fiber-Type Composition by Sponging miR-370-3p Peiwen Zhang 1,2,† , Jingjing Du 1,2,†, Xinyu Guo 1,2, Shuang Wu 1,2, Jin He 1,2 , Xinrong Li 1,2, Linyuan Shen 1,2 , Lei Chen 1,2, Bohong Li 1, Jingjun Zhang 1, Yuhao Xie 1, Lili Niu 1,2, Dongmei Jiang 1,2, Xuewei Li 1,2, Shunhua Zhang 1,2 and Li Zhu 1,2,* 1 College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China; [email protected] (P.Z.); [email protected] (J.D.); [email protected] (X.G.); [email protected] (S.W.); [email protected] (J.H.); [email protected] (X.L.); [email protected] (L.S.); [email protected] (L.C.); [email protected] (B.L.); [email protected] (J.Z.); [email protected] (Y.X.); [email protected] (L.N.); [email protected] (D.J.); [email protected] (X.L.); [email protected] (S.Z.) 2 Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China * Correspondence: [email protected] † These authors contributed equally to this work. Abstract: The development of skeletal muscle is a highly ordered and complex biological process. Increasing evidence has shown that noncoding RNAs, especially long-noncoding RNAs (lncRNAs) and microRNAs, play a vital role in the development of myogenic processes.
    [Show full text]
  • Associated Signaling Complex and Virus-Triggered IRF3 and NF-Κb Activation
    WDR5 is essential for assembly of the VISA- associated signaling complex and virus-triggered IRF3 and NF-κB activation Yan-Yi Wang, Li-Juan Liu, Bo Zhong, Tian-Tian Liu, Ying Li, Yan Yang, Yong Ran, Shu Li, Po Tien, and Hong-Bing Shu1 College of Life Sciences, Modern Virology Center, Wuhan University, Wuhan 430072, China Edited by Philippa Marrack, Howard Hughes Medical Institute/National Jewish, Denver, CO, and approved November 20, 2009 (received for review August 7, 2009) Viral infection causes activation of the transcription factors NF-κBand of VISA in innate antiviral response is also illustrated by the IRF3, which collaborate to induce type I interferons (IFNs) and cellular observation that HCV encoded NS3/4A protease cleaves VISA off antiviral response. The mitochondrial outer membrane protein VISA acts the mitochondria, resulting in abrogation of type I IFN induction as a critical adapter for assembling a virus-induced complex that signals and chronic HCV infection (14). NF-κB and IRF3 activation. Using a biochemical purification approach, On the outer membrane of mitochondria, VISA acts as a cen- we identified the WD repeat protein WDR5 as a VISA-associated protein. tral adapter for assembling a virus-induced complex that activates WDR5 was recruited to VISA in a viral infection dependent manner. distinct signaling pathways leading to IRF3 and NF-κBactivation. Viral infection also caused translocation of WDR5 from the nucleus to VISA is constitutively associated with another mitochondrion- mitochondria. Knockdown of WDR5 impaired the formation of virus- associated adapter protein MITA/STING, which is also essential induced VISA-associated complex.
    [Show full text]
  • The NANOG Transcription Factor Induces Type 2 Deiodinase Expression and Regulates the Intracellular Activation of Thyroid Hormone in Keratinocyte Carcinomas
    Cancers 2020, 12 S1 of S18 Supplementary Materials: The NANOG Transcription Factor Induces Type 2 Deiodinase Expression and Regulates the Intracellular Activation of Thyroid Hormone in Keratinocyte Carcinomas Annarita Nappi, Emery Di Cicco, Caterina Miro, Annunziata Gaetana Cicatiello, Serena Sagliocchi, Giuseppina Mancino, Raffaele Ambrosio, Cristina Luongo, Daniela Di Girolamo, Maria Angela De Stefano, Tommaso Porcelli, Mariano Stornaiuolo and Monica Dentice Figure S1. Strategy for the mutagenesis of Dio2 promoter. (A) Schematic representation of NANOG Binding Site within the Dio2 promoter region. (B) Schematic diagram for site‐directed mutagenesis of NANOG Binding Site on Dio2 promoter region by Recombinant PCR. (C) Representation of the mutated NANOG Binding Site on Dio2 promoter region. (D) Electropherogram of the NANOG Binding Site mutation within the Dio2 promoter. Cancers 2020, 12 S2 of S18 Figure S2. Strategy for the silencing of NANOG expression. (A) Cloning strategies for the generation of NANOG shRNA expression vectors. (B) Electropherograms of the NANOG shRNA sequences cloned into pcDNA3.1 vector. (C) Validation of effective NANOG down-modulation by two different NANOG shRNA vectors was assessed by Western Blot analysis of NANOG expression in BCC cells. (D) Quantification of NANOG protein levels versus Tubulin levels in the same experiment as in C is represented by histograms. Cancers 2020, 12 S3 of S18 Figure S3. The CD34+ cells are characterized by the expression of typical epithelial stemness genes. The mRNA levels of a panel of indicated stemness markers of epidermis were measured by Real Time PCR in the same experiment indicated in figure 3F and G. Cancers 2020, 12 S4 of S18 Figure S4.
    [Show full text]
  • Genetic Factors in Nonsyndromic Orofacial Clefts
    Published online: 2021-02-12 THIEME Review Article 101 Genetic Factors in Nonsyndromic Orofacial Clefts Mahamad Irfanulla Khan1 Prashanth C.S.2 Narasimha Murthy Srinath3 1 Department of Orthodontics & Dentofacial Orthopedics, The Oxford Address for correspondence Mahamad Irfanulla Khan, BDS, MDS, Dental College, Bangalore, Karnataka, India Department of Orthodontics & Dentofacial Orthopedics, The Oxford 2 Department of Orthodontics & Dentofacial Orthopedics, DAPM R.V. Dental College, Bangalore, Karnataka 560068, India Dental College, Bangalore, Karnataka, India (e-mail: [email protected]). 3 Department of Oral & Maxillofacial Surgery, Krishnadevaraya College of Dental Sciences, Bangalore, Karnataka, India Global Med Genet 2020;7:101–108. Abstract Orofacial clefts (OFCs) are the most common congenital birth defects in humans and immediately recognized at birth. The etiology remains complex and poorly understood and seems to result from multiple genetic and environmental factors along with gene– environment interactions. It can be classified into syndromic (30%) and nonsyndromic (70%) clefts. Nonsyndromic OFCs include clefts without any additional physical or Keywords cognitive deficits. Recently, various genetic approaches, such as genome-wide associ- ► orofacial clefts ation studies (GWAS), candidate gene association studies, and linkage analysis, have ► nonsyndromic identified multiple genes involved in the etiology of OFCs. ► genetics This article provides an insight into the multiple genes involved in the etiology of OFCs. ► gene mutation Identification of specific genetic causes of clefts helps in a better understanding of the ► genome-wide molecular pathogenesis of OFC. In the near future, it helps to provide a more accurate association study diagnosis, genetic counseling, personalized medicine for better clinical care, and ► linkage analysis prevention of OFCs.
    [Show full text]
  • Tumor and Stem Cell Biology Research
    Published OnlineFirst September 17, 2012; DOI: 10.1158/0008-5472.CAN-12-0614 Cancer Tumor and Stem Cell Biology Research OTX2 Represses Myogenic and Neuronal Differentiation in Medulloblastoma Cells Ren-Yuan Bai1, Verena Staedtke1, Hart G. Lidov3, Charles G. Eberhart2, and Gregory J. Riggins1 Abstract The brain development transcription factor OTX2 is overexpressed and/or genomically amplified in most medulloblastomas, but the mechanistic basis for its contributions in this setting are not understood. In this study, we identified OTX2 as a transcriptional repressor and a gatekeeper of myogenic and neuronal differentiation in medulloblastoma cells. OTX2 binds to the MyoD1 core enhancer through its homeobox domain, and the remarkable repressor activity exhibited by the homeobox domain renders OTX2 transcrip- tionally repressive. RNA interference–mediated attenuation of OTX2 expression triggered myogenic and neuronal differentiation in vitro and prolonged the survival in an orthotopic medulloblastoma mouse model. Conversely, inducing myogenic conversion of medulloblastoma cells led to the loss of OTX2 expression. In medullomyoblastoma, a medulloblastoma subtype containing muscle elements, myogenic cells share cytogenetic signatures with the primitive tumor cells and OTX2 expression was lost in the differentiated myogenic cells. Thus, OTX2 functions via its homeobox domain as a suppressor of differentiation, and the loss of OTX2 expression is linked to the myogenesis in medullomyoblastoma. Together, our findings illustrate the origin of muscle cells in medullomyoblastomas and the oncogenic mechanism of OTX2 as a repressor of diverse differentiating potential. Cancer Res; 72(22); 1–14. Ó2012 AACR. Introduction is featured with rhabdomyoblastic elements immunoreactive Medulloblastoma, the most common type of pediatric brain to desmin, myoglobin, and fast myosin (3).
    [Show full text]
  • Identification of Shared and Unique Gene Families Associated with Oral
    International Journal of Oral Science (2017) 9, 104–109 OPEN www.nature.com/ijos ORIGINAL ARTICLE Identification of shared and unique gene families associated with oral clefts Noriko Funato and Masataka Nakamura Oral clefts, the most frequent congenital birth defects in humans, are multifactorial disorders caused by genetic and environmental factors. Epidemiological studies point to different etiologies underlying the oral cleft phenotypes, cleft lip (CL), CL and/or palate (CL/P) and cleft palate (CP). More than 350 genes have syndromic and/or nonsyndromic oral cleft associations in humans. Although genes related to genetic disorders associated with oral cleft phenotypes are known, a gap between detecting these associations and interpretation of their biological importance has remained. Here, using a gene ontology analysis approach, we grouped these candidate genes on the basis of different functional categories to gain insight into the genetic etiology of oral clefts. We identified different genetic profiles and found correlations between the functions of gene products and oral cleft phenotypes. Our results indicate inherent differences in the genetic etiologies that underlie oral cleft phenotypes and support epidemiological evidence that genes associated with CL/P are both developmentally and genetically different from CP only, incomplete CP, and submucous CP. The epidemiological differences among cleft phenotypes may reflect differences in the underlying genetic causes. Understanding the different causative etiologies of oral clefts is
    [Show full text]
  • Genomic Analysis of Estrogen Cascade Reveals Histone Variant H2A.Z Associated with Breast Cancer Progression
    Molecular Systems Biology 4; Article number 188; doi:10.1038/msb.2008.25 Citation: Molecular Systems Biology 4:188 & 2008 EMBO and Nature Publishing Group All rights reserved 1744-4292/08 www.molecularsystemsbiology.com Genomic analysis of estrogen cascade reveals histone variant H2A.Z associated with breast cancer progression Sujun Hua1,2,3,9, Caleb B Kallen4,9, Ruby Dhar1,2, Maria T Baquero5, Christopher E Mason6, Beth A Russell1,2,6, Parantu K Shah1,2, Jiang Liu1,2, Andrey Khramtsov7, Maria S Tretiakova8, Thomas N Krausz8, Olufunmilayo I Olopade7, David L Rimm5 and Kevin P White1,2,* 1 Joint Institute for Genomics and Systems Biology, The University of Chicago and Argonne National Laboratory, Chicago, IL, USA, 2 Department of Human Genetics, The University of Chicago, Chicago, IL, USA, 3 Interdepartmental Program in Computational Biology and Bioinformatics, Yale University, New Haven, CT, USA, 4 Department of Gynecology and Obstetrics, Emory University, Atlanta, GA, USA, 5 Department of Pathology, Yale University, New Haven, CT, USA, 6 Department of Genetics, Yale University, New Haven, CT, USA, 7 Center for Clinical Cancer Genetics, University of Chicago Medical Center, Chicago, IL, USA and 8 Department of Pathology, The University of Chicago Hospitals, Chicago, IL, USA 9 These authors contributed equally to this work * Corresponding author. Department of Human Genetics, The University of Chicago, 1639 Pierce Drive, WMB 4211, Chicago, IL 60637, USA. Tel.: þ 1 773 834 8259; Fax: þ 1 773 834 0505; E-mail: [email protected] Received 7.9.07; accepted 13.3.08 We demonstrate an integrated approach to the study of a transcriptional regulatory cascade involved in the progression of breast cancer and we identify a protein associated with disease progression.
    [Show full text]