nach oben

Medical Microbiology and Immunology

Erschienen in:

01.06.2015 | Original Investigation

The proteome of human cytomegalovirus virions and dense bodies is conserved across different strains

verfasst von: Nicole Büscher, Christina Paulus, Michael Nevels, Stefan Tenzer, Bodo Plachter

Erschienen in: Medical Microbiology and Immunology | Ausgabe 3/2015

Einloggen, um Zugang zu erhalten

Abstract

The morphogenesis of human cytomegalovirus (HCMV) particles is incompletely understood. Analysis of the protein composition of HCMV virions and subviral dense bodies (DBs) by mass spectrometry provides valuable information to increase our knowledge about viral morphogenesis. Here we addressed the viral proteome of virions and DBs from two fibroblast-passaged isolates and the widely used endotheliotropic TB4-BAC40 strain of HCMV. The results show a striking concordance of the particle proteomes of different strains. One surprising finding was that only low levels of gpUL128-131A were found in TB40-BAC4 virions. These three proteins, together with gH and gL, form a protein complex that is critical for the endothelial cell tropism of that strain. This indicates that either few molecules of that complex per virion or a small fraction of pentamer-positive virions suffice to retain the tropism. Furthermore, using a pp65-deficient variant of TB40-BAC4, we confirm our previous finding that the major tegument protein serves as a scaffold to support the upload of a fraction of the outer tegument proteins into particles. The results demonstrate that HCMV particle morphogenesis is an orchestrated process that leads to the formation of particles with a largely strain-independent protein composition.

Viswanathan K, Früh K (2007) Viral proteomics: global evaluation of viruses and their interaction with the host. Expert Rev Proteomics 4:815–829CrossRefPubMed

Maxwell KL, Frappier L (2007) Viral proteomics. Microbiol Mol Biol Rev 71:398–411CrossRefPubMedCentralPubMed

Varnum SM, Streblow DN, Monroe ME, Smith P, Auberry KJ, Pasa-Tolic L, Wang D, Camp DG, Rodland K, Wiley S, Britt W, Shenk T, Smith RD, Nelson JA (2004) Identification of proteins in human cytomegalovirus (HCMV) particles: the HCMV proteome. J Virol 78:10960–10966CrossRefPubMedCentralPubMed

Caposio P, Streblow DN, Nelson JA (2013) Cytomegalovirus proteomics. In: Reddehase MJ (ed) Cytomegaloviruses From Molecular Pathogenesis to Intervention. Caister Academic Press, Norfolk, UK, pp 86–108

Reyda S, Tenzer S, Navarro P, Gebauer W, Saur M, Krauter S, Büscher N, Plachter B (2014) The tegument protein pp65 of human cytomegalovirus acts as an optional scaffold protein that optimizes protein uploading into viral particles. J Virol 88:9633–9646CrossRefPubMedCentralPubMed

Reyda S, Büscher N, Tenzer S, Plachter B (2014) Proteomic analyses of human cytomegalovirus strain AD169 derivatives reveal highly conserved patterns of viral and cellular proteins in infected fibroblasts. Viruses 6:172–188CrossRefPubMedCentralPubMed

Dolan A, Cunningham C, Hector RD, Hassan-Walker AF, Lee L, Addison C, Dargan DJ, McGeoch DJ, Gatherer D, Emery VC, Griffiths PD, Sinzger C, McSharry BP, Wilkinson GW, Davison AJ (2004) Genetic content of wild-type human cytomegalovirus. J Gen Virol 85:1301–1312CrossRefPubMed

Dargan DJ, Douglas E, Cunningham C, Jamieson F, Stanton RJ, Baluchova K, McSharry BP, Tomasec P, Emery VC, Percivalle E, Sarasini A, Gerna G, Wilkinson GW, Davison AJ (2010) Sequential mutations associated with adaptation of human cytomegalovirus to growth in cell culture. J Gen Virol 91:1535–1546CrossRefPubMedCentralPubMed

Cunningham C, Gatherer D, Hilfrich B, Baluchova K, Dargan DJ, Thomson M, Griffiths PD, Wilkinson GW, Schulz TF, Davison AJ (2010) Sequences of complete human cytomegalovirus genomes from infected cell cultures and clinical specimens. J Gen Virol 91:605–615CrossRefPubMedCentralPubMed

10.

Murphy E, Yu D, Grimwood J, Schmutz J, Dickson M, Jarvis MA, Hahn G, Nelson JA, Myers RM, Shenk TE (2003) Coding potential of laboratory and clinical strains of human cytomegalovirus. Proc Natl Acad Sci USA 100:14976–14981CrossRefPubMedCentralPubMed

11.

Dunn W, Chou C, Li H, Hai R, Patterson D, Stolc V, Zhu H, Liu F (2003) Functional profiling of a human cytomegalovirus genome. Proc Natl Acad Sci USA 100:14223–14228CrossRefPubMedCentralPubMed

12.

Murphy E, Shenk T (2008) Human cytomegalovirus genome. Curr Top Microbiol Immunol 325:1–19PubMed

13.

Cha TA, Tom E, Kemble GW, Duke GM, Mocarski ES, Spaete RR (1996) Human cytomegalovirus clinical isolates carry at least 19 genes not found in laboratory strains. J Virol 70:78–83PubMedCentralPubMed

14.

Wang D, Shenk T (2005) Human cytomegalovirus virion protein complex required for epithelial and endothelial cell tropism. Proc Natl Acad Sci USA 102:18153–18158CrossRefPubMedCentralPubMed

15.

Wang D, Shenk T (2005) Human cytomegalovirus UL131 open reading frame is required for epithelial cell tropism. J Virol 79:10330–10338CrossRefPubMedCentralPubMed

16.

Gerna G, Percivalle E, Lilleri D, Lozza L, Fornara C, Hahn G, Baldanti F, Revello MG (2005) Dendritic-cell infection by human cytomegalovirus is restricted to strains carrying functional UL131-128 genes and mediates efficient viral antigen presentation to CD8+ T cells. J Gen Virol 86:275–284CrossRefPubMed

17.

Hahn G, Revello MG, Patrone M, Percivalle E, Campanini G, Sarasini A, Wagner M, Gallina A, Milanesi G, Koszinowski U, Baldanti F, Gerna G (2004) Human cytomegalovirus UL131-128 genes are indispensable for virus growth in endothelial cells and virus transfer to leukocytes. J Virol 78:10023–10033CrossRefPubMedCentralPubMed

18.

Webel R, Hakki M, Prichard MN, Rawlinson WD, Marschall M, Chou S (2014) Differential properties of cytomegalovirus pUL97 kinase isoforms affect viral replication and maribavir susceptibility. J Virol 88:4776–4785CrossRefPubMedCentralPubMed

19.

Herget T, Freitag M, Morbitzer M, Kupfer R, Stamminger T, Marschall M (2004) Novel chemical class of pUL97 protein kinase-specific inhibitors with strong anticytomegaloviral activity. Antimicrob Agents Chemother 48:4154–4162CrossRefPubMedCentralPubMed

20.

Sinzger C, Schmidt K, Knapp J, Kahl M, Beck R, Waldman J, Hebart H, Einsele H, Jahn G (1999) Modification of human cytomegalovirus tropism through propagation in vitro is associated with changes in the viral genome. J Gen Virol 80(Pt 11):2867–2877PubMed

21.

Sinzger C, Hahn G, Digel M, Katona R, Sampaio KL, Messerle M, Hengel H, Koszinowski U, Brune W, Adler B (2008) Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E. J Gen Virol 89:359–368CrossRefPubMed

22.

Zalckvar E, Paulus C, Tillo D, Asbach-Nitzsche A, Lubling Y, Winterling C, Strieder N, Mücke K, Goodrum F, Segal E, Nevels M (2013) Nucleosome maps of the human cytomegalovirus genome reveal a temporal switch in chromatin organization linked to a major IE protein. Proc Natl Acad Sci USA 110:13126–13131CrossRefPubMedCentralPubMed

23.

Mücke K, Paulus C, Bernhardt K, Gerrer K, Schön K, Fink A, Sauer EM, Asbach-Nitzsche A, Harwardt T, Kieninger B, Kremer W, Kalbitzer HR, Nevels M (2014) Human cytomegalovirus major immediate early 1 protein targets host chromosomes by docking to the acidic pocket on the nucleosome surface. J Virol 88:1228–1248CrossRefPubMedCentralPubMed

24.

Tischer BK, Smith GA, Osterrieder N (2010) En passant mutagenesis: a two step markerless red recombination system. Methods Mol Biol 634:421–430CrossRefPubMed

25.

Tischer BK, von EJ, Kaufer B, Osterrieder N (2006) Two-step red-mediated recombination for versatile high-efficiency markerless DNA manipulation in Escherichia coli. Biotechniques 40:191–197CrossRefPubMed

26.

Chevillotte M, Landwehr S, Linta L, Frascaroli G, Luske A, Buser C, Mertens T, von Einem J (2009) Major tegument protein pp65 of human cytomegalovirus is required for the incorporation of pUL69 and pUL97 into the virus particle and for viral growth in macrophages. J Virol 83:2480–2490CrossRefPubMedCentralPubMed

27.

Krauss S, Kaps J, Czech N, Paulus C, Nevels M (2009) Physical requirements and functional consequences of complex formation between the cytomegalovirus IE1 protein and human STAT2. J Virol 83:12854–12870CrossRefPubMedCentralPubMed

28.

Distler U, Kuharev J, Navarro P, Levin Y, Schild H, Tenzer S (2014) Drift time-specific collision energies enable deep-coverage data-independent acquisition proteomics. Nat Methods 11:167–170CrossRefPubMed

29.

Yu X, Shah S, Lee M, Dai W, Lo P, Britt W, Zhu H, Liu F, Zhou ZH (2011) Biochemical and structural characterization of the capsid-bound tegument proteins of human cytomegalovirus. J Struct Biol 174:451–460CrossRefPubMedCentralPubMed

30.

Klages S, Rüger B, Jahn G (1989) Multiplicity dependent expression of the predominant phosphoprotein pp65 of human cytomegalovirus. Virus Res 12:159–168CrossRefPubMed

31.

Pari GS (2008) Nuts and bolts of human cytomegalovirus lytic DNA replication. Curr Top Microbiol Immunol 325:153–166PubMed

32.

Adler B, Sinzger C (2013) Cytomegalovirus interstrain variance in cell type tropism. In: Reddehase MJ (ed) Cytomegaloviruses. Caister Academic Press, Norfolk, pp 297–329

33.

Zhou M, Yu Q, Wechsler A, Ryckman BJ (2013) Comparative analysis of gO isoforms reveals that strains of human cytomegalovirus differ in the ratio of gH/gL/gO and gH/gL/UL128-131 in the virion envelope. J Virol 87:9680–9690CrossRefPubMedCentralPubMed

34.

Ryckman BJ, Chase MC, Johnson DC (2010) Human cytomegalovirus TR strain glycoprotein O acts as a chaperone promoting gH/gL incorporation into virions but is not present in virions. J Virol 84:2597–2609CrossRefPubMedCentralPubMed

35.

Scrivano L, Sinzger C, Nitschko H, Koszinowski UH, Adler B (2011) HCMV spread and cell tropism are determined by distinct virus populations. PLoS Pathog 7:e1001256CrossRefPubMedCentralPubMed

36.

Murrell I, Tomasec P, Wilkie GS, Dargan DJ, Davison AJ, Stanton RJ (2013) Impact of sequence variation in the UL128 locus on production of human cytomegalovirus in fibroblast and epithelial cells. J Virol 87:10489–10500CrossRefPubMedCentralPubMed

37.

Plotkin SA, Plachter B (2013) Cytomegalovirus Vaccine: On the Way to the Future? In: Reddehase MJ (ed) Cytomegaloviruses: From Molecular Pathogenesis to Intervention. Caister Academic Press, Norfolk, U.K., pp 424–449

38.

Kabanova A, Perez L, Lilleri D, Marcandalli J, Agatic G, Becattini S, Preite S, Fuschillo D, Percivalle E, Sallusto F, Gerna G, Corti D, Lanzavecchia A (2014) Antibody-driven design of a human cytomegalovirus gHgLpUL128L subunit vaccine that selectively elicits potent neutralizing antibodies. Proc Natl Acad Sci USA 111:17965–17970CrossRefPubMedCentralPubMed

39.

Lilleri D, Kabanova A, Revello MG, Percivalle E, Sarasini A, Genini E, Sallusto F, Lanzavecchia A, Corti D, Gerna G (2013) Fetal human cytomegalovirus transmission correlates with delayed maternal antibodies to gH/gL/pUL128-130-131 complex during primary infection. PLoS ONE 8:e59863CrossRefPubMedCentralPubMed

40.

Lilleri D, Kabanova A, Lanzavecchia A, Gerna G (2012) Antibodies against neutralization epitopes of human cytomegalovirus gH/gL/pUL128-130-131 complex and virus spreading may correlate with virus control in vivo. J Clin Immunol 32:1324–1331CrossRefPubMed

41.

Macagno A, Bernasconi NL, Vanzetta F, Dander E, Sarasini A, Revello MG, Gerna G, Sallusto F, Lanzavecchia A (2010) Isolation of human monoclonal antibodies that potently neutralize human cytomegalovirus infection by targeting different epitopes on the gH/gL/UL128-131A complex. J Virol 84:1005–1013CrossRefPubMedCentralPubMed

42.

Saccoccio FM, Sauer AL, Cui X, Armstrong AE, Habib ESE, Johnson DC, Ryckman BJ, Klingelhutz AJ, Adler SP, McVoy MA (2011) Peptides from cytomegalovirus UL130 and UL131 proteins induce high titer antibodies that block viral entry into mucosal epithelial cells. Vaccine 29:2705–2711CrossRefPubMedCentralPubMed

43.

Freed DC, Tang Q, Tang A, Li F, He X, Huang Z, Meng W, Xia L, Finnefrock AC, Durr E, Espeseth AS, Casimiro DR, Zhang N, Shiver JW, Wang D, An Z, Fu TM (2013) Pentameric complex of viral glycoprotein H is the primary target for potent neutralization by a human cytomegalovirus vaccine. Proc Natl Acad Sci USA 110:E4997–E5005CrossRefPubMedCentralPubMed

44.

Wussow F, Chiuppesi F, Martinez J, Campo J, Johnson E, Flechsig C, Newell M, Tran E, Ortiz J, La RC, Herrmann A, Longmate J, Chakraborty R, Barry PA, Diamond DJ (2014) Human cytomegalovirus vaccine based on the envelope gH/gL pentamer complex. PLoS Pathog 10:e1004524CrossRefPubMedCentralPubMed

45.

Wen Y, Monroe J, Linton C, Archer J, Beard CW, Barnett SW, Palladino G, Mason PW, Carfi A, Lilja AE (2014) Human cytomegalovirus gH/gL/UL128/UL130/UL131A complex elicits potently neutralizing antibodies in mice. Vaccine 32:3796–3804CrossRefPubMed

46.

Mocarski ES, Shenk T, Griffiths PD, Pass RF (2013) Cytomegaloviruses. In: Knipe DM, Howley PM (eds) Fields virology. Wolters Kluwer Lippincott Williams & Wilkins, Philadelphia, pp 1960–2014

Titel: The proteome of human cytomegalovirus virions and dense bodies is conserved across different strains
verfasst von: Nicole Büscher
Christina Paulus
Michael Nevels
Stefan Tenzer
Bodo Plachter
Publikationsdatum: 01.06.2015
Verlag: Springer Berlin Heidelberg
Erschienen in: Medical Microbiology and Immunology / Ausgabe 3/2015
Print ISSN: 0300-8584
Elektronische ISSN: 1432-1831
DOI: https://doi.org/10.1007/s00430-015-0397-y

Leitlinien kompakt für die Innere Medizin

Mit medbee Pocketcards sicher entscheiden.

^{Seit 2022 gehört die medbee GmbH zum Springer Medizin Verlag}

Kostenlos registrieren

Neu im Fachgebiet Innere Medizin

Notfall-TEP der Hüfte ist auch bei 90-Jährigen machbar

26.04.2024 Hüft-TEP Nachrichten

Ob bei einer Notfalloperation nach Schenkelhalsfraktur eine Hemiarthroplastik oder eine totale Endoprothese (TEP) eingebaut wird, sollte nicht allein vom Alter der Patientinnen und Patienten abhängen. Auch über 90-Jährige können von der TEP profitieren.

Niedriger diastolischer Blutdruck erhöht Risiko für schwere kardiovaskuläre Komplikationen

25.04.2024 Hypotonie Nachrichten

Wenn unter einer medikamentösen Hochdrucktherapie der diastolische Blutdruck in den Keller geht, steigt das Risiko für schwere kardiovaskuläre Ereignisse: Darauf deutet eine Sekundäranalyse der SPRINT-Studie hin.

Bei schweren Reaktionen auf Insektenstiche empfiehlt sich eine spezifische Immuntherapie

25.04.2024 Allergien und Intoleranzreaktionen Nachrichten

Insektenstiche sind bei Erwachsenen die häufigsten Auslöser einer Anaphylaxie. Einen wirksamen Schutz vor schweren anaphylaktischen Reaktionen bietet die allergenspezifische Immuntherapie. Jedoch kommt sie noch viel zu selten zum Einsatz.

Therapiestart mit Blutdrucksenkern erhöht Frakturrisiko

25.04.2024 Hypertonie Nachrichten

Beginnen ältere Männer im Pflegeheim eine Antihypertensiva-Therapie, dann ist die Frakturrate in den folgenden 30 Tagen mehr als verdoppelt. Besonders häufig stürzen Demenzkranke und Männer, die erstmals Blutdrucksenker nehmen. Dafür spricht eine Analyse unter US-Veteranen.

Update Innere Medizin

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.

Newsletter bestellen

Springer Medizin

Abstract

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 3/2015

How understanding immunology contributes to managing CMV disease in immunosuppressed patients: now and in future

Stepwise adaptation of murine cytomegalovirus to cells of a foreign host for identification of host range determinants

An endocytic YXXΦ (YRRF) cargo sorting motif in the cytoplasmic tail of murine cytomegalovirus AP2 ‘adapter adapter’ protein m04/gp34 antagonizes virus evasion of natural killer cells

Sleepless latency of human cytomegalovirus

Controversies in the natural history of congenital human cytomegalovirus infection: the paradox of infection and disease in offspring of women with immunity prior to pregnancy