DOCSLIB.ORG

IFIT2 Restriction of WNV Infection Through Ikkɛ Phosphorylation of STAT1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
IFIT2 Restriction of WNV Infection Through Ikkɛ Phosphorylation of STAT1 JBC Papers in Press. Published on November 7, 2011 as Manuscript M111.285205 The latest version is at http://www.jbc.org/cgi/doi/10.1074/jbc.M111.285205 IFIT2 restriction of WNV infection through IKKɛ phosphorylation of STAT1 IKKɛ, STAT1, and IFIT2 define a novel innate immune effector pathway against West Nile virus infection Olivia Perwitasari1,2, Hyelim Cho3, Michael S. Diamond3,4,5, and Michael Gale, Jr.1,2,* 1Department of Immunology, 2Molecular and Cellular Biology Program, University of Washington School of Medicine, Seattle, WA 98195, 3Department of Molecular Microbiology, 4Pathology and Immunology, 5Medicine, Washington University School of Medicine, St. Louis, MO 63110. Running title: IFIT2 restriction of WNV infection through IKKɛ phosphorylation of STAT1 *Address correspondence to: Dr. Michael Gale, Jr., University of Washington School of Medicine, Department of Immunology, Box 357650, Seattle, WA 98195. Phone: (206) 685-7953. Fax: (206) 643- 1013. E-mail: [email protected] Downloaded from Keywords: Interferon, STAT1, IKKɛ, West Nile virus Background: IFN activates JAK-STAT signaling, expression. Biochemical analyses show that where STAT1 phosphorylation is crucial for ISG STAT1 tyrosine dephosphorylation and CRM1- www.jbc.org induction and expression of IFIT2 to limit West mediated STAT1 nuclear-cytoplasmic shuttling Nile virus infection. are required for STAT1 S708 phosphorylation. Results: IKKɛ mediates STAT1 serine 708 When compared to wt mice, WNV infected phosphorylation exclusive of tyrosine IKKɛ-/- mice exhibit enhanced kinetics of virus at NORTHWESTERN UNIV LIBRARY, on December 9, 2011 phosphorylation but dependent on nuclear export dissemination and increased pathogenesis and ISG synthesis. concomitant with loss of STAT1 S708 Conclusion: IKKɛ-mediated STAT1 S708 phosphorylation and IFIT2 expression. Our phosphorylation is crucial for IFIT2 expression to results define an IFN-induced IKKɛ signaling control WNV. pathway of specific STAT1 phosphorylation Significance: We define a novel anti-WNV innate and IFIT2 expression that imparts innate immune effector pathway. antiviral immunity to restrict WNV infection and control viral pathogenesis. SUMMARY West Nile virus is an emerging virus whose West Nile virus (WNV) is an emerging virulence is dependent upon viral evasion of flavivirus that has recently spread into the Western interferon (IFN) and innate immune defenses. hemisphere from points of origin within Asia, The actions of IFN-stimulated genes (ISGs) Africa, or the Middle East (1;2). Infection by impart control of virus infection but the specific WNV is now a leading cause of arboviral ISGs and regulatory pathways that restrict encephalitis and imparts 4% overall case fatality WNV are not defined. Here we show that IKKɛ frequency in the USA (CDC website, http:// phosphorylation of STAT1 at serine 708 (S708) www.cdc.gov/ncidod/dvbid/westnile/index.htm). drives IFIT2 expression to mediate anti-WNV Typically maintained within avian reservoirs, effector function of IFN. WNV infection was WNV is spread to other vertebrates, including enhanced in cells from IKKɛ-/- or IFIT2-/- humans as dead-end hosts, through mosquito bite. mice. In IKKɛ-/- cells the loss of IFN-induced WNV circulates as 2 major lineages and minor IFIT2 expression was linked to lack of STAT1 clades, with specific clades of lineage 1 phosphorylation on S708 but not Y701 nor representing the emergent and virulent strain in the S727. STAT1 S708 phosphorylation occurs North America and elsewhere while lineage 2 independently of IRF-3 but requires signaling strains are typically endemic to Africa and Asia through the IFN-α/β receptor as a late event in and are not known to cause disease in humans the IFN-induced innate immune response that (1;3-6). WNV infection is controlled in part coincides with IKKɛ-responsive ISGs through type-I interferon (IFN) immune defenses 1 Copyright 2011 by The American Society for Biochemistry and Molecular Biology, Inc. IFIT2 restriction of WNV infection through IKKɛ phosphorylation of STAT1 (7;8). IFN actions comprise a major component of modification of the 5’ nontranslated region of the the innate immune response to virus infection, viral RNA mediated by the methyltransferase which serves to restrict virus replication and activity of the viral NS5 protein (23). These spread in part through the actions of interferon- observations define IFIT2 as a critical host factor stimulated genes (ISGs). WNV suppression of IFN of IFN action and WNV restriction, and signaling is linked to virus dissemination, underscore the IFIT2/WNV interaction as a critical neuroinvasion, and virulence of emergent lineage virus/host interface governing innate antiviral 1 strains (8;9). immunity and infection outcome. WNV acutely induces IFN-β expression from IFIT2 is expressed after virus infection infected cells upon engagement of RNA products directly upon IRF-3 activation as well as upon IFN including viral RNA by the RIG-I-like receptors signaling, owing to the presence of both IRF-3 and (RLRs), RIG-I and MDA5 (10-12). The RLRs ISGF3 binding sites in the Ifit2 promoter (24;29- signal the downstream activation of interferon 31). However, STAT1-/- mice failed to induce Downloaded from regulatory factor (IRF)-3 and NF-κB transcription IFIT2 expression in the CNS following LCMV factors through the actions of the IKK-related and WNV-infection in vivo (32). Importantly, kinases (Tank binding kinase 1 (TBK1) and IFN-induced expression of murine IFIT2 is inhibitor of κB kinase epsilon (IKKɛ). As a result, dependent upon STAT1 S708 phosphorylation, as IRF-3 and NF-κB activation drive the expression recently described by TenOever, et al. (21). In this www.jbc.org of IFN-β, other proinflammatory cytokines, respect, Ifit2 is among a set of ISGs, including chemokines, and direct IRF-3-target genes that Adar1, Mx1, and Oas1b and others, whose confer antiviral and immune-activating functions promoters lack purine-rich region upstream of at NORTHWESTERN UNIV LIBRARY, on December 9, 2011 (13;14). Secreted IFN-β drives the innate immune their ISRE which otherwise serves as STAT2 response characterized by ISG expression. This binding sites, wherein STAT1 S708 process is triggered upon IFN-β binding to the phosphorylation is thought to increase the affinity interferon receptor (IFNAR) to induce receptor for STAT1 binding sites to confer gene expression dimerization, autophosphorylation of receptor- by ISGF3 in the absence of the STAT2 binding associated kinases Tyk2 and JAK1 leading to site (21). Notably, STAT1 S708 phosphorylation tyrosine phosphorylation of signal transducer and is induced after IFN treatment of cells. However, activator of transcription (STAT)1 at residue the temporal relationship of S708 phosphorylation Y701, phosphorylation of STAT2, and assembly to other STAT1 phosphorylation sites during IFN- of the STAT1/STAT2/IRF9 ISGF3 complex (15- stimulation, innate immune responses, or WNV 18). ISGF3 function is further augmented or infection is not known. We therefore conducted sustained by STAT1 serine phosphorylation at the current study to define the cell signaling residues S727 and S708 (19-21). ISGF3 pathway and STAT1 phosphorylation interactions translocates to the nucleus and triggers the that drive IFN-induced IFIT2 expression for the transcription of hundreds of ISGs (13;15). ISG restriction of WNV infection. Our results reveal an products serve as immunomodulators and IKKɛ-dependent pathway of STAT1 S708 restriction factors against virus infection (22-24). phosphorylation whose activation requires IFN Among the ISGs IFN-induced protein with signaling for ISG expression and that plays a key tetratricopeptide repeats (IFIT)2, also known as role in the temporal regulation of STAT1 ISG54 has been identified as an ISG restriction phosphorylation and the expression of IFIT2 factor of WNV (23). IFIT2 belongs to the IFIT crucial to the control of WNV infection and gene family whose members function to restrict immunity. virus infection through alteration of cellular protein synthesis (reviewed in (25)), and IFIT2 EXPERIMENTAL PROCEDURES mediates these action by inhibiting eIF3 function Cell culture, interferons, viruses – Parental in translation initiation (26-28). Our recent study wildtype (wt) 2fTGH fibrosarcoma cells, U3A revealed that in the absence of IFN, ectopic IFIT2 (2fTGH-derived mutant cells, deficient in STAT1) expression can impose a blockade that ultimately and U5A (IFRNAR-deficient, provided by Dr. restricts WNV replication. However, emergent George Stark, Cleveland Clinic), immortalized WNV can evade IFIT2 restriction through 2’-O human PH5CH8 hepatocytes (provided by Dr. 2 IFIT2 restriction of WNV infection through IKKɛ phosphorylation of STAT1 Nobuyuki Kato, Okayama University, Japan), and G-3’ (forward) and 5’-CAACTCAGTCTTGATC human embryonic kidney HEK293 cells were TCTCCAGTTCCTTTAGGGCCATC-3’ (reverse) grown in Dulbecco's modified Eagle medium for Y701E. (DMEM) supplemented with 10% fetal bovine pIFIT2-Luc were gifts from and Kineta, Inc. serum (FBS), 2 mM L-glutamine, 1 mM sodium (Seattle); pISG15-Luc and pIFN-β-Luc were pyruvate, antibiotic-antimycotic solution, and 1x described previously (34). pADAR1-CAT was a nonessential amino acids (complete DMEM). gift from Dr. Charles Samuel (UCSD) and Primary mouse embryonic fibroblasts (MEFs) pADAR1-Luc was generated by subcloning into were isolated from IKKɛ-/-, IRF-3-/-, IFNAR-/-, pGL3 Luciferase reporter vector. Transfections IFIT2-/-, and age-matched wt control
Load more

Recommended publications
  • Secretion and LPS-Induced Endotoxin Shock Α Lipopolysaccharide
    IFIT2 Is an Effector Protein of Type I IFN− Mediated Amplification of Lipopolysaccharide (LPS)-Induced TNF- α Secretion and LPS-Induced Endotoxin Shock This information is current as of September 27, 2021. Alexandra Siegfried, Susanne Berchtold, Birgit Manncke, Eva Deuschle, Julia Reber, Thomas Ott, Michaela Weber, Ulrich Kalinke, Markus J. Hofer, Bastian Hatesuer, Klaus Schughart, Valérie Gailus-Durner, Helmut Fuchs, Martin Hrabe de Angelis, Friedemann Weber, Mathias W. Hornef, Ingo B. Autenrieth and Erwin Bohn Downloaded from J Immunol published online 6 September 2013 http://www.jimmunol.org/content/early/2013/09/06/jimmun ol.1203305 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2013/09/06/jimmunol.120330 Material 5.DC1 Why The JI? Submit online. by guest on September 27, 2021 • 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 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 © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published September 6, 2013, doi:10.4049/jimmunol.1203305 The Journal of Immunology IFIT2 Is an Effector Protein of Type I IFN–Mediated Amplification of Lipopolysaccharide (LPS)-Induced TNF-a Secretion and LPS-Induced Endotoxin Shock Alexandra Siegfried,*,1 Susanne Berchtold,*,1 Birgit Manncke,* Eva Deuschle,* Julia Reber,* Thomas Ott,† Michaela Weber,‡ Ulrich Kalinke,x Markus J.
    [Show full text]
  • Small Nucleolar Rnas Determine Resistance to Doxorubicin in Human Osteosarcoma
    International Journal of Molecular Sciences Article Small Nucleolar RNAs Determine Resistance to Doxorubicin in Human Osteosarcoma Martina Godel 1, Deborah Morena 1, Preeta Ananthanarayanan 1, Ilaria Buondonno 1, Giulio Ferrero 2,3 , Claudia M. Hattinger 4, Federica Di Nicolantonio 1,5 , Massimo Serra 4 , 1 2 1, , 1, , Riccardo Taulli , Francesca Cordero , Chiara Riganti * y and Joanna Kopecka * y 1 Department of Oncology, University of Torino, 1026 Torino, Italy; [email protected] (M.G.); [email protected] (D.M.); [email protected] (P.A.); [email protected] (I.B.); [email protected] (F.D.N.); [email protected] (R.T.) 2 Department of Computer Science, University of Torino, 10149 Torino, Italy; [email protected] (G.F.); [email protected] (F.C.) 3 Department of Clinical and Biological Sciences, University of Torino, 10043 Orbassano, Italy 4 Laboratory of Experimental Oncology, Pharmacogenomics and Pharmacogenetics Research Unit, IRCCS Istituto Ortopedico Rizzoli, 40136 Bologna, Italy; [email protected] (C.M.H.); [email protected] (M.S.) 5 Candiolo Cancer Institute, FPO–IRCCS, 10060 Candiolo, Italy * Correspondence: [email protected] (C.R.); [email protected] (J.K.); Tel.: +39-0116705857 (C.R.); +39-0116705849 (J.K.) These authors equally contributed to this work. y Received: 31 May 2020; Accepted: 21 June 2020; Published: 24 June 2020 Abstract: Doxorubicin (Dox) is one of the most important first-line drugs used in osteosarcoma therapy. Multiple and not fully clarified mechanisms, however, determine resistance to Dox. With the aim of identifying new markers associated with Dox-resistance, we found a global up-regulation of small nucleolar RNAs (snoRNAs) in human Dox-resistant osteosarcoma cells.
    [Show full text]
  • Microarray Analysis of Novel Genes Involved in HSV- 2 Infection
    Microarray analysis of novel genes involved in HSV- 2 infection Hao Zhang Nanjing University of Chinese Medicine Tao Liu ( [email protected] ) Nanjing University of Chinese Medicine https://orcid.org/0000-0002-7654-2995 Research Article Keywords: HSV-2 infection,Microarray analysis,Histospecic gene expression Posted Date: May 12th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-517057/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/19 Abstract Background: Herpes simplex virus type 2 infects the body and becomes an incurable and recurring disease. The pathogenesis of HSV-2 infection is not completely clear. Methods: We analyze the GSE18527 dataset in the GEO database in this paper to obtain distinctively displayed genes(DDGs)in the total sequential RNA of the biopsies of normal and lesioned skin groups, healed skin and lesioned skin groups of genital herpes patients, respectively.The related data of 3 cases of normal skin group, 4 cases of lesioned group and 6 cases of healed group were analyzed.The histospecic gene analysis , functional enrichment and protein interaction network analysis of the differential genes were also performed, and the critical components were selected. Results: 40 up-regulated genes and 43 down-regulated genes were isolated by differential performance assay. Histospecic gene analysis of DDGs suggested that the most abundant system for gene expression was the skin, immune system and the nervous system.Through the construction of core gene combinations, protein interaction network analysis and selection of histospecic distribution genes, 17 associated genes were selected CXCL10,MX1,ISG15,IFIT1,IFIT3,IFIT2,OASL,ISG20,RSAD2,GBP1,IFI44L,DDX58,USP18,CXCL11,GBP5,GBP4 and CXCL9.The above genes are mainly located in the skin, immune system, nervous system and reproductive system.
    [Show full text]
  • Rnaseq Reveals the Contribution of Interferon Stimulated Genes to the Increased Host Defense and Decreased PPR Viral Replication in Cattle
    viruses Article RNAseq Reveals the Contribution of Interferon Stimulated Genes to the Increased Host Defense and Decreased PPR Viral Replication in Cattle 1, 1, Krishnaswamy Gopalan Tirumurugaan y , Rahul Mohanchandra Pawar y, Gopal Dhinakar Raj 2,* , Arthanari Thangavelu 3, John A. Hammond 4 and Satya Parida 4,* 1 Department of Animal Biotechnology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600007, India; [email protected] (K.G.T.); [email protected] (R.M.P.) 2 Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600051, India 3 Department of Veterinary Microbiology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600007, India; [email protected] 4 The Pirbright Institute, Ash Road, Pirbright, Surrey GU24 0NF, UK; [email protected] * Correspondence: [email protected] (G.D.R.); [email protected] (S.P.) These authors contributed equally. y Received: 7 March 2020; Accepted: 16 April 2020; Published: 20 April 2020 Abstract: Peste des petits ruminants virus (PPRV) is known to replicate in a wide variety of ruminants causing very species-specific clinical symptoms. Small ruminants (goats and sheep) are susceptible to disease while domesticated cattle and buffalo are dead-end hosts and do not display clinical symptoms. Understanding the host factors that influence differential pathogenesis and disease susceptibility could help the development of better diagnostics and control measures. To study this, we generated transcriptome data from goat and cattle peripheral blood mononuclear cells (PBMC) experimentally infected with PPRV in-vitro. After identifying differentially expressed genes, we further analyzed these immune related pathway genes using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) and selected candidate genes were validated using in-vitro experiments.
    [Show full text]
  • And Chemoresistant MDA-MB-231 Human Breast Cancer Cells Through Up-Regulation of IFIT2
    nutrients Article Baicalein Suppresses Stem Cell-Like Characteristics in Radio- and Chemoresistant MDA-MB-231 Human Breast Cancer Cells through Up-Regulation of IFIT2 So Yae Koh 1, Jeong Yong Moon 2, Tatsuya Unno 2,3 and Somi Kim Cho 1,2,3,* 1 Interdisciplinary Graduate Program in Advanced Convergence Technology and Science, Jeju National University, Jeju 63243, Korea; [email protected] 2 Subtropical/Tropical Organism Gene Bank, Jeju National University, Jeju 63243, Korea; [email protected] (J.Y.M.); [email protected] (T.U.) 3 Faculty of Biotechnology, College of Applied Life Sciences, SARI, Jeju National University, Jeju 63243, Korea * Correspondence: [email protected]; Tel.: +82-64-754-3348 Received: 18 February 2019; Accepted: 11 March 2019; Published: 14 March 2019 Abstract: Resistance to both chemotherapy and radiation therapy is frequent in triple-negative breast cancer (TNBC) patients. We established treatment-resistant TNBC MDA-MB-231/IR cells by irradiating the parental MDA-MB-231 cells 25 times with 2 Gy irradiation and investigated the molecular mechanisms of acquired resistance. The resistant MDA-MB-231/IR cells were enhanced in migration, invasion, and stem cell-like characteristics. Pathway analysis by the Database for Annotation, Visualization and Integrated Discovery revealed that the NF-κB pathway, TNF signaling pathway, and Toll-like receptor pathway were enriched in MDA-MB-231/IR cells. Among 77 differentially expressed genes revealed by transcriptome analysis, 12 genes involved in drug and radiation resistance, including interferon-induced protein with tetratricopeptide repeats 2 (IFIT2), were identified. We found that baicalein effectively reversed the expression of IFIT2, which is reported to be associated with metastasis, recurrence, and poor prognosis in TNBC patients.
    [Show full text]
  • Conventional and Unconventional Mechanisms for Capping Viral Mrna
    REVIEWS Conventional and unconventional mechanisms for capping viral mRNA Etienne Decroly1, François Ferron1, Julien Lescar1,2 and Bruno Canard1 Abstract | In the eukaryotic cell, capping of mRNA 5′ ends is an essential structural modification that allows efficient mRNA translation, directs pre-mRNA splicing and mRNA export from the nucleus, limits mRNA degradation by cellular 5′–3′ exonucleases and allows recognition of foreign RNAs (including viral transcripts) as ‘non-self’. However, viruses have evolved mechanisms to protect their RNA 5′ ends with either a covalently attached peptide or a cap moiety (7‑methyl-Gppp, in which p is a phosphate group) that is indistinguishable from cellular mRNA cap structures. Viral RNA caps can be stolen from cellular mRNAs or synthesized using either a host- or virus-encoded capping apparatus, and these capping assemblies exhibit a wide diversity in organization, structure and mechanism. Here, we review the strategies used by viruses of eukaryotic cells to produce functional mRNA 5′-caps and escape innate immunity. Pre-mRNA splicing The cap structure found at the 5′ end of eukaryotic reactions involved in the viral RNA-capping process, A post-transcriptional mRNAs consists of a 7‑methylguanosine (m7G) moi‑ and the specific cellular factors that trigger a response modification of pre-mRNA, in ety linked to the first nucleotide of the transcript via a from the innate immune system. which introns are excised and 5′–5′ triphosphate bridge1 (FIG. 1a). The cap has several exons are joined in order to form a translationally important biological roles, such as protecting mRNA Capping, decapping and turnover of host RNA functional, mature mRNA.
    [Show full text]
  • Prolactin-Releasing Peptide Differentially Regulates Gene Transcriptomic Profiles in Mouse Bone Marrow-Derived Macrophages
    International Journal of Molecular Sciences Article Prolactin-Releasing Peptide Differentially Regulates Gene Transcriptomic Profiles in Mouse Bone Marrow-Derived Macrophages Yulong Sun 1,2,* , Zhuo Zuo 1,2 and Yuanyuan Kuang 1,2 1 School of Life Sciences, Northwestern Polytechnical University, Xi’an 710072, China; [email protected] (Z.Z.); [email protected] (Y.K.) 2 Key Laboratory for Space Biosciences & Biotechnology, Institute of Special Environmental Biophysics, School of Life Sciences, Northwestern Polytechnical University, Xi’an 710072, China * Correspondence: [email protected]; Tel.: +86-29-8846-0332 Abstract: Prolactin-releasing Peptide (PrRP) is a neuropeptide whose receptor is GPR10. Recently, the regulatory role of PrRP in the neuroendocrine field has attracted increasing attention. However, the influence of PrRP on macrophages, the critical housekeeper in the neuroendocrine field, has not yet been fully elucidated. Here, we investigated the effect of PrRP on the transcriptome of mouse bone marrow-derived macrophages (BMDMs) with RNA sequencing, bioinformatics, and molecular simulation. BMDMs were exposed to PrRP (18 h) and were subjected to RNA sequencing. Differentially expressed genes (DEGs) were acquired, followed by GO, KEGG, and PPI analysis. Eight qPCR-validated DEGs were chosen as hub genes. Next, the three-dimensional structures of the proteins encoded by these hub genes were modeled by Rosetta and Modeller, followed by molecular dynamics simulation by the Gromacs program. Finally, the binding modes between PrRP Citation: Sun, Y.; Zuo, Z.; Kuang, Y. and hub proteins were investigated with the Rosetta program. PrRP showed no noticeable effect on Prolactin-Releasing Peptide the morphology of macrophages.
    [Show full text]
  • Full Text (PDF)
    medRxiv preprint doi: https://doi.org/10.1101/2020.12.29.20248986; this version posted January 4, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . Insights into the molecular mechanism of anticancer drug ruxolitinib repurposable in COVID-19 therapy Manisha Mandal Department of Physiology, MGM Medical College, Kishanganj-855107, India Email: [email protected], ORCID: https://orcid.org/0000-0002-9562-5534 Shyamapada Mandal* Department of Zoology, University of Gour Banga, Malda-732103, India Email: [email protected], ORCID: https://orcid.org/0000-0002-9488-3523 *Corresponding author: Email: [email protected]; [email protected] Abstract Due to non-availability of specific therapeutics against COVID-19, repurposing of approved drugs is a reasonable option. Cytokines imbalance in COVID-19 resembles cancer; exploration of anti-inflammatory agents, might reduce COVID-19 mortality. The current study investigates the effect of ruxolitinib treatment in SARS-CoV-2 infected alveolar cells compared to the uninfected one from the GSE5147507 dataset. The protein-protein interaction network, biological process and functional enrichment of differentially expressed genes were studied using STRING App of the Cytoscape software and R programming tools. The present study indicated that ruxolitinib treatment elicited similar response equivalent to that of SARS-CoV-2 uninfected situation by inducing defense response in host against virus infection by RLR and NOD like receptor pathways. Further, the effect of ruxolitinib in SARS- CoV-2 infection was mainly caused by significant suppression of IFIH1, IRF7 and MX1 genes as well as inhibition of DDX58/IFIH1-mediated induction of interferon- I and -II signalling.
    [Show full text]
  • Computational and Transcriptome Analyses Revealed Preferential Induction of Chemotaxis and Lipid Synthesis by SARS-Cov-2
    biology Article Computational and Transcriptome Analyses Revealed Preferential Induction of Chemotaxis and Lipid Synthesis by SARS-CoV-2 Hibah Shaath 1,2 and Nehad M. Alajez 1,2,* 1 College of Health & Life Sciences, Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF), PO Box 34110 Doha, Qatar; [email protected] 2 Cancer Research Center, Qatar Biomedical Research Institute (QBRI), Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF), PO Box 34110 Doha, Qatar * Correspondence: [email protected]; Tel.: +974-4454-7252 Received: 27 July 2020; Accepted: 26 August 2020; Published: 1 September 2020 Abstract: The continuous and rapid emergence of new viral strains calls for a better understanding of the fundamental changes occurring within the host cell upon viral infection. In this study, we analyzed RNA-seq transcriptome data from Calu-3 human lung epithelial cells infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) compared to five other viruses namely, severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East Respiratory Syndrome (SARS-MERS), influenzavirus A (FLUA), influenzavirus B (FLUB), and rhinovirus (RHINO) compared to mock-infected cells and characterized their coding and noncoding RNA transcriptional portraits. The induction of interferon, inflammatory, and immune response was a hallmark of SARS-CoV-2 infection. Comprehensive bioinformatics revealed the activation of immune response and defense response to the virus as a common feature of viral infection. Interestingly however, the degree of functional categories and signaling pathways activation varied among different viruses. Ingenuity pathways analysis highlighted altered conical and casual pathways related to TNF, IL1A, and TLR7, which are seen more predominantly during SARS-CoV-2 infection.
    [Show full text]
  • IFIT2 (F-12): Sc-390724
    SANTA CRUZ BIOTECHNOLOGY, INC. IFIT2 (F-12): sc-390724 BACKGROUND STORAGE The tetratricopeptide repeat (TPR) motif is a degenerate, 34 amino acid Store at 4° C, **DO NOT FREEZE**. Stable for one year from the date of sequence found in many proteins and acts to mediate protein-protein inter- shipment. Non-hazardous. No MSDS required. actions in various pathways. At the sequence level, there can be up to 16 tandem TPR repeats, each of which has a helix-turn-helix shape that stacks DATA on other TPR repeats to achieve ligand binding specificity. IFIT2 (interferon- induced protein with tetratricopeptide repeats 2), also known as G10P2 or IFI54, is a 472 amino acid protein that contains six TPR repeats and may be 116 K – 81 K – involved in the negative regulation of cell growth and proliferation. The gene encoding IFIT2 maps to human chromosome 10, which houses over 1,200 <IFIT2 genes and comprises nearly 4.5% of the human genome. Defects in some of 50 K – the genes that map to chromosome 10 are associated with Charcot-Marie- Tooth disease, Jackson-Weiss syndrome, Usher syndrome, nonsyndromatic deafness, Wolman’s syndrome, Cowden syndrome, multiple endocrine neoplasia type 2 and porphyria. IFIT2 (F-12): sc-390724. Western blot analysisofIFIT2 IFIT2 (F-12): sc-390724. Immunoperoxidase staining of expression in SCC-25 wholecell lysate. formalin fixed, paraffin-embedded human kidney tissue showing cytoplasmic staining of cells in tubules. CHROMOSOMAL LOCATION Genetic locus: IFIT2 (human) mapping to 10q23.31. SELECT PRODUCT CITATIONS 1. Sanpui, P., et al. 2014. Single-walled carbon nanotubes increase pandemic SOURCE influenza A H1N1 virus infectivity of lung epithelial cells.
    [Show full text]
  • Identification and Validation of Key Genes Associated with Systemic Sclerosis-Related Pulmonary Hypertension
    fgene-11-00816 July 23, 2020 Time: 17:24 # 1 ORIGINAL RESEARCH published: 24 July 2020 doi: 10.3389/fgene.2020.00816 Identification and Validation of Key Genes Associated With Systemic Sclerosis-Related Pulmonary Hypertension Ji-Na Zheng1†, Yang Li2†, Yue-Mei Yan1, Hui Shi3, Tian-Tian Zou4,5, Wen-Qi Shao6 and Qiang Wang1* 1 Department of Dermatology, Zhongshan Hospital, Fudan University, Shanghai, China, 2 Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 3 Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China, 4 Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, China, 5 Cancer Metastasis Institute, Fudan University, Shanghai, China, 6 Department of Laboratory Medicine, Zhongshan Hospital, Fudan University, Shanghai, China Systemic sclerosis-associated with pulmonary arterial hypertension (SSc-PAH) is still a major cause of SSc related deaths. Early diagnosis and prompt treatment are crucial to reduce the mortality of patients with SSc-PAH. To screen the candidate biomarkers Edited by: and potential therapeutic targets for SSc-PAH, we analyzed the data set (GSE33463 Juan Caballero, and GSE19617) for confirming key genes in peripheral blood mononuclear cells from Universidad Autónoma de Querétaro, Mexico SSc-PAH patients. A total of 105 SSc patients from gene expression omnibus (GEO) Reviewed by: were included as discovery cohort (n = 69) and duplication cohort (n = 36) for screening Gil Speyer, hub genes by weighted gene co-expression network analysis (WGCNA). Furthermore, Arizona State University, United States Yan Gong, an independent validation cohort (n = 40), including healthy controls, SSc and SSc- Wuhan University, China PAH patients, was used for further validation by quantitative real-time polymerase *Correspondence: chain reaction.
    [Show full text]
  • SAHA Overcomes 5-FU Resistance in IFIT2-Depleted Oral Squamous Cell Carcinoma Cells
    cancers Article SAHA Overcomes 5-FU Resistance in IFIT2-Depleted Oral Squamous Cell Carcinoma Cells Prabha Regmi 1,2, Kuo-Chu Lai 3, Chung-Ji Liu 4 and Te-Chang Lee 2,* 1 Taiwan International Graduate Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei 11529, Taiwan; [email protected] 2 Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan 3 Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan City 33302, Taiwan; [email protected] 4 Department of Oral and Maxillofacial Surgery, Mackay Memorial Hospital, Taipei 10421, Taiwan; [email protected] * Correspondence: [email protected] Received: 30 October 2020; Accepted: 24 November 2020; Published: 26 November 2020 Simple Summary: IFIT2 depletion is associated with increased epithelial-mesenchymal transition and metastasis. The main aim of our study was to understand the link between drug resistance and IFIT2 depletion. In this study, we confirmed resistance to multiple common therapeutic drugs, particularly 5-FU, which showed especially high resistance in IFIT2-depleted cells. Interestingly, combination of SAHA and 5-FU overcame 5-FU resistance in IFIT2-depleted cells. Hence, our findings suggest that IFIT2 expression may be used as a biomarker to decide whether to undergo 5-FU treatment, but also the SAHA and 5-FU combination may be a potential new treatment regimen to augment 5-FU therapy in patients with thymidylate synthase-mediated drug-resistant oral squamous cell carcinoma. Abstract: Interferon-induced protein with tetratricopeptide repeats 2 (IFIT2) is a member of the interferon-stimulated gene family that contains tetratricopeptide repeats (TPRs), which mediate protein–protein interactions in various biological systems.
    [Show full text]