Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Human cytomegalovirus immediate-early-gene expression disrupts embryogenesis in transgenic Drosophila

  • 143 Accesses

  • 9 Citations


Intrauterine infection with human cytomegalovirus (HCMV) is the leading viral cause of birth defects involving the central nervous system. Due to the highly species specific nature of the virus, its course of natural infection cannot be studied in animal models. Here we introduce a novel transgenic Drosophila model system for studying the effects of the major viral regulatory genes, the immediate-early genes, on normal embryonic development. We show that ectopic expression of the immediate-early genes in Drosophila led to increased embryonic lethality manifested in disintegration of the embryos. Further analysis suggested that immediate-early gene expression interfered with adherens junction maintenance, leading to the disruption of embryonic epithelial integrity. Owing to the evolutionary conservation of developmental mechanisms from invertebrates to mammals, we anticipate that the studies in Drosophila will be relevant also to humans and will ultimately provide a versatile system for studying different aspects of viral-host interactions.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7


  1. Aberle H, Bauer A, Stappert J, Kispert A, Kemler R (1997) β-catenin is a target for the ubiquitin-proteasome pathway. EMBO J 16:3797–3804

  2. Adamson AL, Wright N, LaJeunesse DR (2005) Modeling early Epstein-Barr virus infection in Drosophila melanogaster, The BZLF1 Protein. Genetics 171:1125–1135

  3. Battaglia PA, Zito S, Macchini A, Gigliani F (2001) A Drosophila model of HIV-Tat-related pathogenicity. J Cell Sci 114:2787–294

  4. Belvin MP, Anderson KV (1996) A conserved signaling pathway: The Drosophila Toll-Dorsal pathway. Annu Rev Cell Dev Biol 2:393–416

  5. Bissinger AL, Sinzger C, Kaiserling E, Jahn G. (2002) Human cytomegalovirus as a direct pathogen: correlation of multiorgan involvement and cell distribution with clinical and pathological findings in a case of congenital inclusion disease. J Medical Virol 67:200–206

  6. Bonini NM, Fortini ME (2003) Human neurodegenerative disease modeling using Drosophila. Annu Rev Neurosci 26:627–656

  7. Boshart M, Weber F, Jahn G, Dorsch-Hasler K, Fleckenstein B, Schaffner W (1985) A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell 41:521–530

  8. Brand AH, Manoukian AS, Perrimon N. 1994. Ectopic expression in Drosophila. Methods Cell Biol 44:635–654

  9. Cadigan K, Nusse R (1997) Wnt signaling, a common theme in animal development. Genes Dev 11:3286–3305

  10. Campos-Ortega JA, Hartenstein V (1985) The embryonic development of Drosophila melanogaster. Springer Verlag, Berlin

  11. Castillo JP, Kowalik TF (2002) Human cytomegalovirus immediate early proteins and cell growth control. Gene 290:19–34

  12. Castillo JP, Kowalik TF (2004) HCMV infection, modulating the cell cycle and cell death. Int Rev Immunol 23:113–39

  13. Challacombe JF, Rechtsteiner A, Gottardo R, Rocha LM, Browne EP, Shenk T, Altherr MR, Brettin TS (2004) Evaluation of the host transcriptional response to human cytomegalovirus infection. Physiol Genomics 18:51–62

  14. Chambers J, Angulo A, Amaratunga D, Guo H, Jiang Y, Wan JS, Bittener 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

  15. Chen YT, Stewart DB, Nelson WJ (1999) Coupling assembly of the E-cadherin/beta-catenin complex to efficient endoplasmic reticulum exit and basal-lateral membrane targeting of E-cadherin in polarized MDCK cells. J Cell Biol 144:687–699

  16. Conacci-Sorrell M, Zhurinsky J, Ben-Ze’ev A (2002) The cadherin-catenin adhesion system in signaling and cancer. J Clin Invest 109:987–991

  17. Danilkovitch-Miagkova A, Miagkov A, Skeel A, Nakaigawa N, Zbar B, Leonard EJ (2001) Oncogenic mutants of RON and MET receptor tyrosine kinases cause activation of the beta-catenin pathway. Mol Cell Biol 21:5857–5868

  18. DeMeritt IB, Milford LE, Yurochko AD (2004). Activation of the NF-κB pathway in human Cytomegalovirus-infected cells is necessary for efficient transactivation of the major immediate-early promoter. J Virol 78:4498–4507

  19. Deshpande G, Stukey J, Schedl P (1995) scute (sis-b) function in Drosophila sex determination. Mol Cell Biol 15:4430–4440

  20. Foecking MK, Hofstetter H (1986) Powerful and versatile enhancer/promoter unit for mammalian expression vectors. Gene 45:101–105

  21. Gaytant MA, Steegers EA, Semmekrot BA, Merkus HM, Galama JM (2002) Congenital cytomegalovirus infection, review of the epidemiology and outcome. Obstet Gynecol Surv 57:245–256

  22. Ghazal P, Nelson JA (1993) Transcription factors and viral regulatory proteins as potential mediators of human cytomegalovirus pathogenesis. In: Becker Y, Darai G, Huang ES (eds) Molecular aspects of human cytomegalovirus diseases. Springer-Verlag, Berlin, pp 360–383

  23. Hafen E (2004) Cancer, type 2 diabetes, and ageing, news from flies and worms. Swiss Med Wkly 134:711–719

  24. Hamada F, Bienz M (2002) A Drosophila APC tumour suppressor homologue functions in cellular adhesion. Nat Cell Biol 4:208–213

  25. Henderson BR, Fagotto F (2002) The ins and outs of APC and β-Catenin nuclear transport. EMBO reports 3:834–839

  26. Ho M (1991) Cytomegalovirus, biology and infection, 2nd edn. Plenum Publishing, New York

  27. Jamora C, Fuchs E (2002) Intercellular adhesion, signalling and the cytoskeleton. Nat Cell Biol 4:E101–E108

  28. Kalejta RF, Shenk T (2002) Manipulation of the cell cycle by human cytomegalovirus. Front Biosci 7:295–306

  29. Kang DE, Soriano S, Xia X, Eberhart CG, De Strooper B, Zheng H, Koo EH (2002) Presenilin couples the paired phosphorylation of β-catenin independent of axin, implications for β-catenin activation in tumorigenesis. Cell 110:751–762

  30. Koedood M, Fichtel A, Meier P, Mitchell PJ (1995) Human cytomegalovirus HCMV immediate-early enhancer/promoter specificity during embryogenesis defines target tissues of congenital HCMV infection. J Virol 69:2194–2207

  31. Landolfo S, Gariglio M, Gribaudo G, Lembo D (2003) The human cytomegalovirus. Pharmacol Ther 98:269–297

  32. Lasko P (2002) Diabetic flies? Using Drosophila melanogaster to understand the causes of monogenic and genetically complex diseases. Clin Genet 62:358–367

  33. Li R-Y, Baba S, Kosugi I, Arai Y, Kawasaki H, Shinmura Y, Sakakibara S-I, Okano H, Tsutsui Y (2001) Activation of murine cytomegalovirus immediate-early promoter in cerebral ventricular zone and glial progenitor cells in transgenic mice. Glia 35:41–52

  34. Lilien J, Balsamo J (2005) The regulation of cadherin-mediated adhesion by tyrosine phosphorylation/dephosphorylation of β-catenin. Curr Opin Cell Biol 17:459–465

  35. Macara IG (2004) Parsing the polarity code. Nat Rev Mol Cell Biol 5:220–231

  36. McGuirore SE, Roman G, Davis RL (2004) Gene expression systems in Drosophila, a synthesis of time and space. Trends Genet 20:384–391

  37. Michelson S, Alcami J, Kim S-J, Danielpour D, Bachelerie F, Picard L, Bessia C, Paya C, Virelizier J-L (1994) Human Cytomegalovirus infection induces transcription and secretion of transforming growth factor beta-1. J Virol 68:5730–5737

  38. Müller HA, Wieschaus E (1996) Armadillo, bazooka, and stardust are critical for early stages in formation of the zonula adherens and maintenance of the polarized blastoderm epithelium in Drosophila. J Cell Biol 134:149–163

  39. Müller HAJ, Bossinger O (2003) Molecular networks controlling epithelial cell polarity in development. Mech Dev 120:1231–1256

  40. Pajovic S, Wong EL, Black AR, Azizkhan CJ (1997) Identification of a viral kinase that phosphorylates specific E2Fs and pocket proteins. Mol Cell Biol 17:6459–6464

  41. Park CS, Kim SI, Lee MS, Youn C, Kim DJ, Jho E, Song WK (2004) Modulation of β-Catenin phosphorylation/degradation by cyclin-dependent kinase 2. J Biol Chem 19:19592–19599

  42. Pass RF (2001) Cytomegalovirus. In: Knipe DM, Howley PM (eds) Fields virology, 4th edn. Lippincott Wiliams and Wilkins, Philadelphia, pp 2675–2706

  43. Pass RF, Stagno S, Myers GJ, Alford CA (1980) Outcome of symptomatic congenital cytomegalovirus infection, results of long-term longitudinal follow-up. Pediatrics 66:758–762

  44. Peifer M, Sweeton D, Casey M, Wieschaus E (1994) Wingless signal and Zeste-white 3 kinase trigger opposing changes in the intracellular distribution of Armadillo. Development 120:369–380

  45. Piedra J, Miravet S, Castaño J, Pálmer HG, Heisterkamp N, García de Herreros A, Duñach M (2003) p120 Catenin-associated Fer and Fyn tyrosine kinases regulate beta-catenin Tyr-142 phosphorylation and beta-catenin-alpha-catenin Interaction. Mol Cell Biol 23:2287–2297

  46. Pirrotta V (1988) Vectors for P-mediated transformation of Drosophila. In: Rodriguez RL, Denhardt DT (eds) Vectors, a survey of molecular cloning vectors and their uses. Butterworths, Boston, pp 437–456

  47. Plachter B, Sinzger C, Jahn G (1996) Cell types involved in replication and distribution of human cytomegalovirus. Adv Virus Res 46:195–261

  48. Polakis P (2002) Casein kinase 1, A Wnt’er of disconnect. Curr Biol 12:R499–R501

  49. Schleiss MR (2002) Animal models of congenital cytomegalovirus infection, an overview of progress in the characterization of guinea pig cytomegalovirus GPCMV. J Clin Virol 25:37–49

  50. Scholz M, Blaheta RA, Vogel J-U, Doerr HW, Cinat J Jr, (1999) Cytomegalovirus-induced trans endothelial cell migration; a closer look at intercellular communication mechanisms. Intervirol 42:350–356

  51. Shulman JM, Shulman LM, Weiner WJ, Feany MB (2003) From fruit fly to bedside, translating lessons from Drosophila models of neurodegenerative disease. Curr Opin Neurol 16:443–449

  52. Sinclair J, Baillie J, Bryant L, Caswell R (2000) Human cytomegalovirus mediates cell cycle progression through G1 into early S phase in terminally differentiated cells. J Gen Virol 8:1553–1565

  53. Spector DH. (1996) Activation and regulation of human cytomegalovirus early genes. Intervirol 39:361–377

  54. Spradling AC, Rubin GM. (1982) Transposition of cloned P elements into Drosophila germ line chromosomes. Science 218:343–347

  55. Staczek J (1990) Animal cytomegaloviruses. Microbiol Rev 54:247–265

  56. Stoker AW (2005) Protein tyrosine phosphatases and signaling. J Endocrinol 185:19–33

  57. Tarantal AF, Salamat MS, Britt WJ, Luciw PA, Hendrickx AG, Barry PA (1998) Neuropathogenesis induced by rhesus cytomegalovirus in fetal rhesus monkeys (Macaca mulatta). J Infect Dis 177:446–450

  58. Tepass U, Tanentzapf G, Ward R, Fehon R (2001) Epithelial cell polarity and cell junctions in Drosophila. Annu Rev Genet 35:747–784

  59. Thummel CS, Pirrotta V (1992) New CaspeR P-element vectors. Dros Info Serv 71:150

  60. Tzou P, De Gregorio E, Lemaitre B (2002) How Drosophila combats microbial infection, a model to study innate immunity and host-pathogen interactions. Curr Opin Microbiol 5:102–110

  61. Vančíková Z, Dvořák P (2001) Cytomegalovirus infection in immunocompetent and immunocompromised individuals Current Drug Targets-Immune. Endocr Metab Disord 1:179–187

  62. Vogelmann R, Nguyen-tat M-D, Gieh K, Adler G, Wedlich D, Menke A (2005) TGF-β induced down regulation of E-cadherin-based cell-cell adhesion depends on PI3-kinase and PTEN. J Cell Sci 118:4901–4912

  63. Wang Z, Lindquist S (1998) Developmentally regulated nuclear transport of transcription factors in Drosophila embryos enables the heat shock response. Development 125:4841–4850

  64. Xu G, Craig AW, Greer P, Miller M, Anastasiadis PZ, Lilien J, Balsamo J (2004) Continuous association of cadherin with β-catenin requires the non-recpetor tyrosine-kinase Fer. J Cell Sci 117:3207–3219

  65. Yamaguchi M, Hirose F, Inoue YH, Ohno K, Yoshida H, Hayashi Y, Deak P, Matsukage A (2004) Genetic link between p53 and genes required for formation of the zonula adherens junction. Cancer Sci 95:436–441

  66. Yu Y, Alwine JC (2002) Human cytomegalovirus major immediate-early proteins and simian virus 40 large T antigen can inhibit apoptosis through activation of the phosphatidylinositide 3’-OH kinase pathway and the cellular kinase Akt. J Vir 76:3731–3738

Download references


We thank Hava Haikin, Virology Department at Soroka Mediacl Center, Israel, for HCMV infected cells; Alain Debec, Gavdos group, France, for the GFP trap line; Dr. John Belote, Syracuse University, USA, for transgenic line harboring an Hsp70-GFP construct; Inbal Bulukan for help with embryo injections. This work was supported in part by a grant from the ‘Israel Ministry of Health’ to F.S.N-S and by an institutional award to F.S.N-S.

Author information

Correspondence to Shira Neuman-Silberberg.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Steinberg, R., Shemer-Avni, Y., Adler, N. et al. Human cytomegalovirus immediate-early-gene expression disrupts embryogenesis in transgenic Drosophila . Transgenic Res 17, 105–119 (2008). https://doi.org/10.1007/s11248-007-9136-5

Download citation


  • HCMV
  • Immediate early
  • Birth defects
  • Drosophila model
  • Adhesion