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Die Familie Der Herpesviridae Herpesviridae

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Die Familie Der Herpesviridae Herpesviridae Die Familie der Herpesviridae Herpesviridae Unterfamile Genus Vertreter (Mensch) α-Herpesvirinae Simplexvirus Humanes Herpesvirus 1 (Herpes- simplex-Virus-1) Humanes Herpesvirus 2 (Herpes- simplex-Virus-2) Varicellovirus Humanes Herpesvirus 3 (Varicella Zoster Virus) β-Herpesvirinae Cytomegalovirus Humanes Herpesvirus 5 Muromegalovirus Kein Roseolovirus Humanes Herpesvirus 6,7 γ-Herpesvirinae Lymphocryptovirus Humanes Herpesvirus 4 (Epstein Barr Virus) Rhadinovirus Humanes Herpesvirus 8 (= Kaposi sarcoma herpes virus) Molekulare Virologie Ruth Brack-Werner; SS 2008 Herpesviridae: Gemeinsame, charakteristische MerkmaleHerpesviridae . • Partikel enthalten Faktoren, die u.a. die virale Transkription aktivieren und die in die zelluläre Genexpression eingreifen; • Zusammenbau der Viruskapside und Virus-Genomsynthese im Zellkern; • Virusvermehrung = Zerstörung der Wirtszelle; • Latenz; • Kodieren mehrere Enzyme/Proteine für die Genomreplikation (z.B. DNA Polymerase, Korrekturenzyme und Enzyme für die Bereitstellung von Nukleotid-Bausteinen ); • Genome linear, dsDNA, bis zu 230 kB lang. Molekulare Virologie Ruth Brack-Werner; SS 2008 Herpesviridae: Merkmale der einzelnen Unterfamilien Herpesviridae Zell-Spektrum Zell-Spektrum Zell-Spektrum http://www.vu-wien.ac.at/i123/SPEZVIR/HERPESGEN1.HTML Molekulare Virologie Ruth Brack-Werner; SS 2008 Struktur von Herpesviren Herpesviridae Lipidhülle; Tegument Grösse: 180-200 nM Durchmesser; Lipidmembran Schematische Darstellung (Beispiel: Elektronen Mikroskopische HCMV); Aufnahme http://www.biografix.de/ http://web.uct.ac.za/depts/mmi/stannard/emimages.html Molekulare Virologie Ruth Brack-Werner; SS 2008 Herpesvirus Genome Herpesviridae http://www-micro.msb.le.ac.uk/3035/3035pics/Herpes1.gif Molekulare Virologie Ruth Brack-Werner; SS 2008 Isomere des Cytomegalovirus Genoms Herpesviridae Molekulare Virologie Aus”Molekulare Virologie”, Modrow, Falk, Truyen, 2.Auflage, 2003, Spektrum Ruth Brack-Werner; SS 2008 Akademischer Verlag, Kap. 19l, Fig.19.24B Herpesvirus Genome liegen als Episom in infizierter Zelle vor Herpesviridae Gene für: DNA Synthese Kapsid Membranproteine LAT Replikationsursprung http://darwin.bio.uci.edu/~faculty/wagner/hsvimg04.jpg Molekulare Virologie Ruth Brack-Werner; SS 2008 Beispiele für Herpesvirus Promotoren Herpesviridae Fast jedes Gen hat seinen eigenen Promotor. Aus Fields Virology 4th edition, 2002, Chapter727, Lippincott, Williams and Wilkins, 2002 Fig. 67-6 Molekulare Virologie Ruth Brack-Werner; SS 2008 Beispiel eines viralen Transkriptionsregulators: HSV VP16 Herpesviridae Latency VP16 /bacteria/hhv1/herpes.html stdgen / gov 500 - 1000 . lanl molecules . stdgen α-genes β-genes γ-genes Immediate early early late + particle:http://www. - Virus Molekulare Virologie Ruth Brack-Werner; SS 2008 „Immediate early“ (α) virale Proteine: Beispiel HSV Herpesviridae Proteine Modifikation/Grösse (kD) Funktionen ICP4 Phosphoryliert; UDP und Aktivierung der Transkription von β und γ Genen; ADP-Reste/ Repression der Transkription von α-Genen (ICP0, 140 ICP4) ICP0 79 Transkriptionsaktivator; wirkt synergistisch mit ICP4; ICP27 Phosphoryliert/ 63 Post-transkriptioneller Regulator; Trägt zur Ausschaltung der Expression zellulärer Proteine bei indem es Splicing inhibiert* Fördert den Export ungespleißter viraler mRNAs; ICP47 Verhindert Präsentation von viralen Antigenen für CD8 Zellen; Molekulare Virologie Ruth Brack-Werner; SS 2008 Replikationsgabel in Säugetierzellen © 2002 by Bruce Alberts, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, and Peter Walter „Early“ Proteine/Enzyme für die virale DNA Replikation (I) Funktion Grösse (kD) /Genort ) HSV CMV EBV HHV-8 Bindet an oriLyt 94/ 70 38-40/ ? UL9 /UL84 BZLF1 DNA Polymerase:Polymerase und 140/ 150/ 110/ Polymerase- Exonukleaseaktvität UL30 UL50 BALF-5 Orf 9 komplex Bindet ds DNA (processivity factor) 62/ 52/ 50 orf59 UL42 UL44 /BMRF1 Bindung von Einzelstrang DNA an 124/ 140/ 138/ 110/ Replikationsgabel UL29 UL57 BALF2 Orf6 5’,-3’Helikase, DNA-Primasekomplex 99/ 115/ -/BSLF1 orf56 UL5 UL105 Helikase- 5’,-3’Helikase, DNA-Primasekomplex 114/ 110/ - 0rf40/41 Primase- /BBLF2/3 Komplex UL52 UL70 Stimuliert Primersynthese 80/UL8 -/UL102? -/BBL4 Orf44 Replikation des Genoms zur Verpackung in Virione: „Rolling circle“ Mechanismus Herpesviridae Aus”An Introduction to Genetic Analysis” Griffiths, Miller, Suzuki, Lewontin, Gelbart, , 7th Ed, 2000, W.H. Freeman and Co, Fig. 8-19, Molekulare Virologie Ruth Brack-Werner; SS 2008 Beteiligung von Herpesvirus Proteinen bei der lytischen Genomreplikation Herpesviridae Aus”Molekulare Virologie”, Modrow, Falk, Truyen, 2.Auflage, 2003, Spektrum Akademischer Verlag, Kap. 19l, Fig.19.25 Molekulare Virologie Ruth Brack-Werner; SS 2008 „Early“ Proteine/Enzyme für die virale DNA Replikation (II) Aktivität Grösse (kD) /Genort ) HSV CMV EBV HHV-8 Thymidinkinase; Phosphoryliert Thymidine und 41/ -/- 70/ andere Nukleoside; UL23 BXLF-1 Orf 21 Ribonukleotidreduktase* Untereinheit (gross) 140/ -/ 85/ Orf61 UL39 UL45 BORF2 Ribonukleotidreduktase* Untereinheit (klein) 38/ ? 34/ Orf60 UL40 BaRF1 Uracilglycosylase: Katalysiert die Freisetzung von 39/ -/ 78-88 orf46 Uracil aus DNA: DNA Reparatur? UL2 UL114 /BKRF3 Alkalische Endo-/Exonuklease: Auflösung von 68/UL12 -/UL98 70/BGLF5 Orf37 Verzweigten Strukturen in der DNA. dUTPase: dUTP -> dUMP zur Verhinderung des 39/ -/ -/BLLF3 orf55 Einbaus von dUTP in die DNA; UL50 UL72 *Katalysiert die Umwandlung von Ribonukleotiden zu Desoxyribonukleotiden; Essentiell für die Replikation in nicht-teilenden Zellen. Exemplarische „Späte“ Proteine Herpesviridae Aktivität Grösse (kD)/Bezeichnung /Genort ) HSV CMV EBV HHV-8 Tegument Protein; aktiviert die Transkription der 54/α-TIF, 71/-/UL82 -/-/BPLF1? “immediate early” Gene ICP25, VP16, UL48 Hauptkapsidprotein 155/VP5/ 153/-/UL86 154/- 153/- UL19 /BCLF1 /orf25 Diverse Membranproteine (Glykosyliert) Molekulare Virologie Ruth Brack-Werner; SS 2008 VP16 funktioniert als Komplex mit zellularen Faktoren Herpesviridae Acidic activation domain HCF-1: human factor C1 Oct-1: Octamer 1 transcription factor Wysocka J and Herr W. 2003.TIBS 28, 294-304. POU domains Molekulare Virologie Ruth Brack-Werner; SS 2008 Replikationszyklus: Eintritt in die Zelle (HSV) Herpesviridae = Entry 3-O-S: 3-O-Sulfotransferase; HS: Heparan sulphate spezifische durch 3-O-S modifizierte HS können als Fusionsrezeptoren dienen. HVEM: Herpes virus entry mediator: Mitglied der TNF-α Rezeptor Familie Nectin 1, 2: Mitglieder der Immunoglobulin superfamilie Spear PG et al. 2004, Molekulare Virologie Ruth Brack-Werner; SS 2008 Heparin Sulphat Herpesviridae Disaccharid bestehend GAG: aus D-glucosamin und N- glycosaminoglycan Acetyl-D-Glucosamin http://www.med.unibs.it/~airc/hspgs.html Negative Ladungen Aus: Molecular Biology of the Cell. 4th ed. Alberts, Bruce; Johnson, Alexander; Lewis, Julian; Raff, Martin; Roberts, Keith; Walter, Peter. New York: Garland Publishing; 2002.Fig. 19-39 Molekulare Virologie Ruth Brack-Werner; SS 2008 Bindung von Herpesviren an Zelloberfläche führt zur Transduktion von Signale (Bsp. HCMV) Herpesviridae Molekulare Virologie Compton T. 2004. TCB. 14, 5-9. Ruth Brack-Werner; SS 2008 Replikationszyklus: HSV I “Immediate Early” “Späte” Ereignisse 12- Ereignisse 1-9: 24-: DNA Replikation: Anheftung des Virus an Bildung von Zelloberfläche; Fusion Konkatameren, die zur der viralen und Transkription der γ - zellulären Membrane; Gene verwendet Eintritt des Kapsids und werden; Synthese von Ausschüttung der Hüllproteinen am ER; Tegumentproteine in die Transport von viralen Zelle; Transport des NC Glykoproteinen in den zur Kernpore und Golgi; Zusammenbau Freisetzung der HSV von Nukleo-Kapsiden DNA in den Kern; (NC) im Kern; Expression der α-Gene Ausknospung von NC in und Synthese der das ER und dann ins Proteine; Zytoplasma; Fusion der zytoplasm. NC mit Golgi-Membran; Erneute Behüllung des Virus; Ausknospung des “Early” Ereignisse 10- behüllten Virus in 11: α-Proteine aktivieren Vesikel und Freisetzung die Synthese der β- der Vesikel durch Proteine; Exozytose Principles of Virology, 2004. Flint SJ, Enquist LW, Racaniello VR,Skalka AM, 2nd edition. ASM Press. Appendix, Fig. 6. Verbreitung von HSV im Wirt Herpesviridae Principles of Virology, 2004. Flint SJ, Enquist LW, Racaniello VR,Skalka AM, Molekulare Virologie Ruth Brack-Werner; SS 2008 2nd edition. ASM Press. Appendix, p.853. HSV-Seroprevalenz Herpesviridae Anteil der Bevölkerung Alter 40% 12-19 > 80% über 60 Molekulare Virologie Ruth Brack-Werner; SS 2008 Weitere HSV-1 und HSV-2 assoziierte Krankheiten Herpesviridae http://pathmicro.med.sc.edu/virol/herpes.htm Principles of Virology, 2004. Flint SJ, Enquist LW, Racaniello VR,Skalka AM, Molekulare Virologie Ruth Brack-Werner; SS 2008 2nd edition. ASM Press. Appendix, p.853. HSV Latenz und Aktivierung Herpesviridae http://pathmicro.med.sc.edu/virol/herpes.htm Molekulare Virologie Ruth Brack-Werner; SS 2008 Reaktivierung von HSV Herpesviridae • Verletzungen; • Schwere Erkrankungen mit Intubationen; • Chirurgische Eingriffe in dem Trigeminus Nerv; • Immunsuppression, z.B. nach Organtransplantationen; Molekulare Virologie Ruth Brack-Werner; SS 2008 Latente Infektion (HSV) Herpesviridae Schritte 1-6: Wie bei produktiver Infektion; Zirkularisierung des viralen Genoms; Snythese der LAT (Latency Associated Transcripts Neuron in Ganglion Principles of Virology, 2004. Flint SJ, Enquist LW, Racaniello VR,Skalka AM, Molekulare Virologie Ruth Brack-Werner; SS 2008 2nd edition. ASM Press. Appendix, Fig. 6. HSV Latenz LAT RNA Herpesviridae • Die LAT RNAs
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