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Transcriptional analysis of human cytomegalovirus and rat cytomegalovirus homologues of the M73/M73.5 spliced gene family

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Abstract

Previous analysis of the M73-to-m74/M75 intergenic region of murine cytomegalovirus (MCMV) identified a family of 3′-co-terminal spliced transcripts that includes M73 and M73.5. The current study investigated whether similar families of spliced genes also exist in the human CMV (HCMV) and rat CMV (RCMV) genomes. Northern blot, RT-PCR and RACE-PCR analysis of HCMV transcripts showed that while mRNAs from HCMV UL73 and a putative UL73.5 homologue were spliced and 3′-co-terminal, they were not 5′-co-terminal. In contrast, the spliced RCMV R73 and R73.5 transcripts were arranged in a similar manner to those in MCMV and found to be both 5′ and 3′-co-terminal. In both the HCMV and RCMV genomes, additional non-coding spliced transcripts were found to originate from these regions. These results highlight that families of spliced transcripts coding for structural glycoproteins are likely to be a conserved feature of this region of betaherpesviral genomes.

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References

  1. Akter P, Cunningham C, McSharry BP, Dolan A, Addison C, Dargan DJ, Hassan-Walker AF, Emery VC, Griffiths PD, Wilkinson GW, Davison AJ (2003) Two novel spliced genes in human cytomegalovirus. J Gen Virol 84:1117–1122

    Article  PubMed  CAS  Google Scholar 

  2. Awasthi S, Isler JA, Alwine JC (2004) Analysis of splice variants of the immediate-early 1 region of human cytomegalovirus. J Virol 78:8191–8200

    Article  PubMed  CAS  Google Scholar 

  3. Brocchieri L, Kledal TN, Karlin S, Mocarski ES (2005) Predicting coding potential from genome sequence: application to betaherpesviruses infecting rats and mice. J Virol 79:7570–7596

    Article  PubMed  CAS  Google Scholar 

  4. Bühler B, Keil GM, Weiland F, Koszinowski UH (1990) Characterization of the murine cytomegalovirus early transcription unit e1 that is induced by immediate-early proteins. J Virol 64:1907–1919

    PubMed  Google Scholar 

  5. Casarosa P, Gruijthuijsen YK, Michel D, Beisser PS, Holl J, Fitzsimons CP, Verzijl D, Bruggeman CA, Mertens T, Leurs R, Vink C, Smit MJ (2003) Constitutive signaling of the human cytomegalovirus-encoded receptor UL33 differs from that of its rat cytomegalovirus homolog R33 by promiscuous activation of G proteins of the Gq, Gi, and Gs classes. J Biol Chem 278:50010–50023

    Article  PubMed  CAS  Google Scholar 

  6. 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 70:78–83

    PubMed  CAS  Google Scholar 

  7. Chambers J, Angulo A, Amaratunga D, Guo H, Jiang Y, Wan JS, Bittner A, Frueh K, Jackson MR, Peterson PA, Erlander MG, Ghazal P (1999) DNA microarrays of the complex human cytomegalovirus genome: profiling kinetic class with drug sensitivity of viral gene expression. J Virol 73:5757–5766

    PubMed  CAS  Google Scholar 

  8. Chee MS, Bankier AT, Beck S, Bohni R, Brown CM, Cerny R, Horsnell T, Hutchison CA, Kouzarides T, Martignetti JA, Preddie E, Satchwell SC, Tomlinson P, Weston KM, Barrell BG (1990) Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol 154:125–169

    PubMed  CAS  Google Scholar 

  9. Ciocco-Schmitt GM, Karabekian Z, Godfrey EW, Stenberg RM, Campbell AE, Kerry JA (2002) Identification and characterization of novel murine cytomegalovirus M112–113 (e1) gene products. Virology 294:199–208

    Article  PubMed  CAS  Google Scholar 

  10. Compton T (2004) Receptors and immune sensors: the complex entry path of human cytomegalovirus. Trends Cell Biol 14:5–8

    Article  PubMed  CAS  Google Scholar 

  11. Davis-Poynter NJ, Lynch DM, Vally H, Shellam GR, Rawlinson WD, Barrell BG, Farrell HE (1997) Identification and characterization of a G protein-coupled receptor homolog encoded by murine cytomegalovirus. J Virol 71:1521–1529

    PubMed  CAS  Google Scholar 

  12. 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 85:1301–1312

    Article  PubMed  CAS  Google Scholar 

  13. Fleming P, Davis-Poynter N, Degli-Esposti M, Densley E, Papadimitriou J, Shellam G, Farrell H (1999) The murine cytomegalovirus chemokine homolog, m131/129, is a determinant of viral pathogenicity. J Virol 73:6800–6809

    PubMed  CAS  Google Scholar 

  14. Frohman MA, Dush MK, Martin GR (1988) Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002

    Article  PubMed  CAS  Google Scholar 

  15. Grey F, Antoniewicz A, Allen E, Saugstad J, McShea A, Carrington JC, Nelson J (2005) Identification and characterization of human cytomegalovirus-encoded microRNAs. J Virol 79:12095–12099

    Article  PubMed  CAS  Google Scholar 

  16. Kaptein SJ, van Cleef KW, Gruijthuijsen YK, Beuken EV, van Buggenhout L, Beisser PS, Stassen FR, Bruggeman CA, Vink C (2004) The r131 gene of rat cytomegalovirus encodes a proinflammatory CC chemokine homolog which is essential for the production of infectious virus in the salivary glands. Virus Genes 29:43–61

    Article  PubMed  CAS  Google Scholar 

  17. Kattenhorn LM, Mills R, Wagner M, Lomsadze A, Makeev V, Borodovsky M, Ploegh HL, Kessler BM (2004) Identification of proteins associated with murine cytomegalovirus virions. J Virol 78:11187–11197

    Article  PubMed  CAS  Google Scholar 

  18. Keil GM, Ebeling-Keil A, Koszinowski UH (1987) Sequence and structural organization of murine cytomegalovirus immediate-early gene 1. J Virol 61:1901–1908

    PubMed  CAS  Google Scholar 

  19. Keil GM, Ebeling-Keil A, Koszinowski UH (1987) Immediate-early genes of murine cytomegalovirus: location, transcripts, and translation products. J Virol 61:526–533

    PubMed  CAS  Google Scholar 

  20. Krmpotić A, Bubić I, Polić B, Lucin P, Jonjić S (2003) Pathogenesis of murine cytomegalovirus infection. Microbes Infect 5:1263–1277

    Article  PubMed  Google Scholar 

  21. Loh LC (1989) Synthesis and processing of a 22–26 K murine cytomegalovirus glycoprotein recognized by a neutralizing monoclonal antibody. Virology 169:474–478

    Article  PubMed  CAS  Google Scholar 

  22. MacDonald MR, Burney MW, Resnick SB, Virgin HW (1999) Spliced mRNA encoding the murine cytomegalovirus chemokine homolog predicts a beta chemokine of novel structure. J Virol 73:3682–3691

    PubMed  CAS  Google Scholar 

  23. Mach M, Kropff B, Dal Monte P, Britt W (2000) Complex formation by human cytomegalovirus glycoproteins M (gpUL100) and N (gpUL73). J Virol 74:11881–11892

    Article  PubMed  CAS  Google Scholar 

  24. Mach M, Kropff B, Kryzaniak M, Britt W (2005) Complex formation by glycoproteins M and N of human cytomegalovirus: structural and functional aspects. J Virol 79:2160–2170

    Article  PubMed  CAS  Google Scholar 

  25. Mount SM (1982) A catalogue of splice junction sequences. Nucleic Acids Res 10:459–472

    Article  PubMed  CAS  Google Scholar 

  26. Murphy E, Rigoutsos I, Shibuya T, Shenk TE (2003) Reevaluation of human cytomegalovirus coding potential. Proc Natl Acad Sci USA 100:13585–13590

    Article  PubMed  CAS  Google Scholar 

  27. 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 100:14976–14981

    Article  PubMed  CAS  Google Scholar 

  28. Pignatelli S, Dal Monte P, Landini MP (2001) gpUL73 (gN) genomic variants of human cytomegalovirus isolates are clustered into four distinct genotypes. J Gen Virol 82:2777–2784

    PubMed  CAS  Google Scholar 

  29. Pignatelli S, Dal Monte P, Rossini G, Chou S, Gojobori T, Hanada K, Guo JJ, Rawlinson W, Britt W, Mach M, Landini MP (2003) Human cytomegalovirus glycoprotein N (gpUL73-gN) genomic variants: identification of a novel subgroup, geographical distribution and evidence of positive selective pressure. J Gen Virol 84:647–655

    Article  PubMed  CAS  Google Scholar 

  30. Rawlinson WD, Farrell HE, Barrell BG (1996) Analysis of the complete DNA sequence of murine cytomegalovirus. J Virol 70:8833–8849

    PubMed  CAS  Google Scholar 

  31. Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Press, Cold Spring Harbor

    Google Scholar 

  32. Sandford GR, Ho K, Burns WH (1993) Characterization of the major locus of Immediate-Early genes of rat cytomegalovirus. J Virol 67:4093–4103

    PubMed  CAS  Google Scholar 

  33. Scalzo AA, Dallas PB, Forbes CA, Mikosza AS, Fleming P, Lathbury LJ, Lyons PA, Laferte S, Craggs MM, Loh LC (2004) The murine cytomegalovirus M73.5 gene, a member of a 3′ co-terminal alternatively spliced gene family, encodes the gp24 virion glycoprotein. Virology 329:234–250

    PubMed  CAS  Google Scholar 

  34. Senapathy P, Shapiro MB, Harris NL (1990) Splice junctions, branch point sites, and exons: sequence statistics, identification, and applications to genome project. Meth Enzymol 183:252–278

    Article  PubMed  CAS  Google Scholar 

  35. Smith LM, Tonkin JN, Lawson MA, Shellam GR (2004) Isolates of cytomegalovirus (CMV) from the black rat Rattus rattus form a distinct group of rat CMV. J Gen Virol 85:1313–1317

    Article  PubMed  CAS  Google Scholar 

  36. Smith LM, McWhorter AR, Masters LL, Shellam GR, Redwood AJ (2008) Laboratory strains of murine cytomegalovirus are genetically similar to but phenotypically distinct from wild strains of virus. J Virol 82:6689–6696

    Article  PubMed  CAS  Google Scholar 

  37. Staprans SI, Spector DH (1986) 2.2-kilobase class of early transcripts encoded by cell-related sequences in human cytomegalovirus strain AD169. J Virol 57:591–602

    PubMed  CAS  Google Scholar 

  38. Stenberg RM, Witte PR, Stinski MF (1985) Multiple spliced and unspliced transcripts from human cytomegalovirus immediate-early region 2 and evidence for a common initiation site within immediate-early region 1. J Virol 56:665–675

    PubMed  CAS  Google Scholar 

  39. Varnum SM, Streblow DN, Monroe ME, Smith P, Auberry KJ, Pasa-Tolic L, Wang D, Camp DG 2nd, 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 78:10960–10966

    Article  PubMed  CAS  Google Scholar 

  40. Vink C, Beuken E, Bruggeman CA (2000) Complete DNA sequence of the rat cytomegalovirus genome. J Virol 74:7656–7665

    Article  PubMed  CAS  Google Scholar 

  41. Wright DA, Spector DH (1989) Posttranscriptional regulation of a class of human cytomegalovirus phosphoproteins encoded by an early transcription unit. J Virol 63:3117–3127

    PubMed  CAS  Google Scholar 

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Acknowledgments

Dr. Helen Farrell and Prof. William Rawlinson are thanked for critically reading the manuscript. This work was supported by grants from the National Health and Medical Research of Australia.

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Authors and Affiliations

  1. Immunology and Virology Program, Centre for Ophthalmology and Visual Science, The University of Western Australia, Nedlands, WA, 6009, Australia

    Anthony A. Scalzo & Catherine A. Forbes

  2. Centre for Experimental Immunology, Lions Eye Institute, 2 Verdun St, Nedlands, WA, 6009, Australia

    Anthony A. Scalzo & Catherine A. Forbes

  3. Discipline of Microbiology, School of Biomedical and Chemical Sciences, University of Western Australia, Nedlands, WA, 6509, Australia

    Lee M. Smith

  4. Department of Biochemistry, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada

    Lambert C. Loh

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  1. Anthony A. Scalzo
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  2. Catherine A. Forbes
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Correspondence to Anthony A. Scalzo.

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Scalzo, A.A., Forbes, C.A., Smith, L.M. et al. Transcriptional analysis of human cytomegalovirus and rat cytomegalovirus homologues of the M73/M73.5 spliced gene family. Arch Virol 154, 65–75 (2009). https://doi.org/10.1007/s00705-008-0274-8

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  • DOI: https://doi.org/10.1007/s00705-008-0274-8

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