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Volume 8, Issue 2

Human cytomegalovirus strain-specific differences in protein expression of type I IFN pathway proteins do not impact virus replication Open Access

Abstract

The type I IFN response is crucial for cells to restrict viral replication during infection. Many viruses, including human cytomegalovirus (HCMV), have evolved mechanisms to antagonize the type I IFN response. We have previously observed an increase in protein expression of certain IFN-stimulated genes when comparing the high-passage HCMV strain AD169 to the low-passage strain HCMV Merlin, suggesting that AD169 is defective in its ability to inhibit type I IFN function. To better understand HCMV interaction with the type I IFN response, we examined expression of cellular and viral proteins expressed in Merlin- and AD169-infected cells associated with IFN production and signalling. HCMV IFN antagonists expressed by both viruses had differences in amino acids throughout their protein sequences, although analysis using AlphaFold revealed that there was likely to be no obvious differences in the overall structure of these proteins. Analysis of quantitative mass spectrometry datasets showed modest differences in the expression of cellular IFN-associated proteins between strains. Contrary to previously reported data, we found no obvious loss of IRF3 expression, though this may be due to experimental differences between studies. These data revealed that multiplicity of infection was an important factor in IRF3 expression. We found little or no statistical difference in the production of IFN- RNA between Merlin- and AD169-infected cells in reverse transcriptase quantitative PCR assays and little or no statistical difference in replication of AD169 and Merlin in virus replication assays. Overall, these data suggest that different strains of HCMV have different, albeit modest, abilities to influence the expression of type I IFN pathway proteins during infection. However, this had no overall impact on the ability of different strains to produce a type I IFN or to replicate.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): alphafold , cytomegalovirus , human and IFN
Funding
This study was supported by the:
  • Medical Research Council (Award MR/W006677/1)
    • Principal Award Recipient: KatieA Latham
  • Deutsche Forschungsgemeinschaft (Award 390874280)
    • Principal Award Recipient: JensBosse
  • Deutsche Forschungsgemeinschaft (Award 453548970)
    • Principal Award Recipient: JensBosse
  • Deutsche Forschungsgemeinschaft (Award 49735088)
    • Principal Award Recipient: JensBosse
© 2026 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2026-02-19
2026-03-12

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References

  1. Marshall JS, Warrington R, Watson W, Kim HL. An introduction to immunology and immunopathology. Allergy Asthma Clin Immunol 2018; 14:49 [View Article]
     [Google Scholar]
  2. Alandijany T. Host intrinsic and innate intracellular immunity during herpes simplex virus type 1 (HSV-1) infection. Front Microbiol 2019; 10:2611 [View Article] [PubMed]
     [Google Scholar]
  3. Wang Y-Q, Zhao X-Y. Human cytomegalovirus primary infection and reactivation: insights from virion-carried molecules. Front Microbiol 2020; 11:1511 [View Article] [PubMed]
     [Google Scholar]
  4. Fletcher-Etherington A, Nobre L, Nightingale K, Antrobus R, Nichols J et al. Human cytomegalovirus protein pUL36: a dual cell death pathway inhibitor. Proc Natl Acad Sci USA 2020; 117:18771–18779 [View Article]
     [Google Scholar]
  5. Dell’Oste V, Biolatti M, Galitska G, Griffante G, Gugliesi F et al. Tuning the orchestra: HCMV vs. innate immunity. Front Microbiol 2020; 11:661 [View Article] [PubMed]
     [Google Scholar]
  6. Rossini G, Cerboni C, Santoni A, Landini MP, Landolfo S et al. Interplay between human cytomegalovirus and intrinsic/innate host responses: a complex bidirectional relationship. Mediators Inflamm 2012; 2012:607276 [View Article] [PubMed]
     [Google Scholar]
  7. Griffante G, Hewelt-Belka W, Albano C, Gugliesi F, Pasquero S et al. IFI16 impacts metabolic reprogramming during human cytomegalovirus infection. mBio 2022; 13:e0043522 [View Article] [PubMed]
     [Google Scholar]
  8. Sapuan S, Theodosiou AA, Strang BL, Heath PT, Jones CE. A systematic review and meta-analysis of the prevalence of human cytomegalovirus shedding in seropositive pregnant women. Rev Med Virol 2022; 32:e2399 [View Article] [PubMed]
     [Google Scholar]
  9. Lee C-H, Grey F. Systems Virology and human cytomegalovirus: using high throughput approaches to identify novel host-virus interactions during lytic infection. Front Cell Infect Microbiol 2020; 10:280 [View Article] [PubMed]
     [Google Scholar]
  10. Chappell JD, Dermody TS. Biology of viruses and viral diseases. Mand Douglas Bennetts Princ Pract Infect Dis 2015 [View Article]
     [Google Scholar]
  11. Marques M, Ferreira AR, Ribeiro D. The interplay between human cytomegalovirus and pathogen recognition receptor signaling. Viruses 2018; 10:514 [View Article] [PubMed]
     [Google Scholar]
  12. Koyama S, Ishii KJ, Coban C, Akira S. Innate immune response to viral infection. Cytokine 2008; 43:336–341 [View Article] [PubMed]
     [Google Scholar]
  13. Lee AJ, Ashkar AA. The dual nature of type I and type II interferons. Front Immunol 2018; 9:2061 [View Article] [PubMed]
     [Google Scholar]
  14. Lin K-M, Nightingale K, Soday L, Antrobus R, Weekes MP. Rapid degradation pathways of host proteins during HCMV infection revealed by quantitative proteomics. Front Cell Infect Microbiol 2020; 10:578259 [View Article] [PubMed]
     [Google Scholar]
  15. Mazewski C, Perez RE, Fish EN, Platanias LC. Type I interferon (IFN)-regulated activation of canonical and non-canonical signaling pathways. Front Immunol 2020; 11:606456 [View Article] [PubMed]
     [Google Scholar]
  16. Zhang B, Goraya MU, Chen N, Xu L, Hong Y et al. Zinc finger CCCH-type antiviral protein 1 restricts the viral replication by positively regulating type I interferon response. Front Microbiol 2020; 11:1912 [View Article] [PubMed]
     [Google Scholar]
  17. Martí-Carreras J, Maes P. Human cytomegalovirus genomics and transcriptomics through the lens of next-generation sequencing: revision and future challenges. Virus Genes 2019; 55:138–164 [View Article] [PubMed]
     [Google Scholar]
  18. Pérez-Carmona N, Martínez-Vicente P, Farré D, Gabaev I, Messerle M et al. A prominent role of the human cytomegalovirus UL8 glycoprotein in restraining proinflammatory cytokine production by myeloid cells at late times during infection. J Virol 2018; 92:e02229-17 [View Article] [PubMed]
     [Google Scholar]
  19. Nobre LV, Nightingale K, Ravenhill BJ, Antrobus R, Soday L et al. Human cytomegalovirus interactome analysis identifies degradation hubs, domain associations and viral protein functions. Elife 2019; 8:e49894 [View Article] [PubMed]
     [Google Scholar]
  20. Goodwin CM, Ciesla JH, Munger J. Who’s driving? Human cytomegalovirus, interferon, and NFκB signaling. Viruses 2018; 10:447 [View Article] [PubMed]
     [Google Scholar]
  21. Griffiths P, Reeves M. Pathogenesis of human cytomegalovirus in the immunocompromised host. Nat Rev Microbiol 2021; 19:759–773 [View Article] [PubMed]
     [Google Scholar]
  22. Nightingale K, Lin K-M, Ravenhill BJ, Davies C, Nobre L et al. High-definition analysis of host protein stability during human cytomegalovirus infection reveals antiviral factors and viral evasion mechanisms. Cell Host Microbe 2018; 24:447–460 [View Article] [PubMed]
     [Google Scholar]
  23. Lista MJ, Witney AA, Nichols J, Davison AJ, Wilson H et al. Strain-dependent restriction of human cytomegalovirus by zinc finger antiviral proteins. J Virol 2023; 97:e0184622 [View Article] [PubMed]
     [Google Scholar]
  24. McSharry BP, Jones CJ, Skinner JW, Kipling D, Wilkinson GWG. Human telomerase reverse transcriptase-immortalized MRC-5 and HCA2 human fibroblasts are fully permissive for human cytomegalovirus. J Gen Virol 2001; 82:855–863 [View Article] [PubMed]
     [Google Scholar]
  25. Sievers F, Higgins DG. Clustal omega for making accurate alignments of many protein sequences. Protein Science 2018; 27:135–145 [View Article]
     [Google Scholar]
  26. Dereeper A, Guignon V, Blanc G, Audic S, Buffet S et al. Phylogeny.fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Res 2008; 36:W465–9 [View Article] [PubMed]
     [Google Scholar]
  27. Soh TK, Ognibene S, Sanders S, Schäper R, Kaufer BB et al. A proteome-wide structural systems approach reveals insights into protein families of all human herpesviruses. Nat Commun 2024; 15:10230 [View Article] [PubMed]
     [Google Scholar]
  28. Weekes MP, Tomasec P, Huttlin EL, Fielding CA, Nusinow D et al. Quantitative temporal viromics: an approach to investigate host-pathogen interaction. Cell 2014; 157:1460–1472 [View Article] [PubMed]
     [Google Scholar]
  29. Choi H jin, Park A, Kang S, Lee E, Lee TA et al. Human cytomegalovirus-encoded US9 targets MAVS and STING signaling to evade type I interferon immune responses. Nat Commun 2018; 9:125 [View Article]
     [Google Scholar]
  30. Mathers C, Schafer X, Martínez-Sobrido L, Munger J. The human cytomegalovirus UL26 protein antagonizes NF-κB activation. J Virol 2014; 88:14289–14300 [View Article] [PubMed]
     [Google Scholar]
  31. Huang Z-F, Zou H-M, Liao B-W, Zhang H-Y, Yang Y et al. Human cytomegalovirus protein UL31 inhibits DNA sensing of cGAS to mediate immune evasion. Cell Host Microbe 2018; 24:69–80 [View Article] [PubMed]
     [Google Scholar]
  32. Fu Y-Z, Su S, Gao Y-Q, Wang P-P, Huang Z-F et al. Human cytomegalovirus tegument protein UL82 inhibits STING-mediated signaling to evade antiviral immunity. Cell Host Microbe 2017; 21:231–243 [View Article] [PubMed]
     [Google Scholar]
  33. Biolatti M, Dell’Oste V, Pautasso S, Gugliesi F, von Einem J et al. Human cytomegalovirus tegument protein pp65 (pUL83) dampens type I interferon production by inactivating the DNA sensor cGAS without affecting STING. J Virol 2018; 92:e01774-17 [View Article] [PubMed]
     [Google Scholar]
  34. Kim J-E, Kim Y-E, Stinski MF, Ahn J-H, Song Y-J. Human cytomegalovirus IE2 86 kDa protein induces STING degradation and inhibits cGAMP-mediated IFN-β induction. Front Microbiol 2017; 8:1854 [View Article] [PubMed]
     [Google Scholar]
  35. Paulus C, Krauss S, Nevels M. A human cytomegalovirus antagonist of type I IFN-dependent signal transducer and activator of transcription signaling. Proc Natl Acad Sci USA 2006; 103:3840–3845 [View Article] [PubMed]
     [Google Scholar]
  36. Gatherer D, Seirafian S, Cunningham C, Holton M, Dargan DJ et al. High-resolution human cytomegalovirus transcriptome. Proc Natl Acad Sci U S A 2011; 108:19755–19760 [View Article] [PubMed]
     [Google Scholar]
  37. Dargan DJ, Jamieson FE, MacLean J, Dolan A, Addison C et al. The published DNA sequence of human cytomegalovirus strain AD169 lacks 929 base pairs affecting genes UL42 and UL43. J Virol 1997; 71:9833–9836 [View Article] [PubMed]
     [Google Scholar]
  38. Sinzger C, Hahn G, Digel M, Katona R, Sampaio KL et al. Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E. J Gen Virol 2008; 89:359–368 [View Article] [PubMed]
     [Google Scholar]
  39. Cunningham C, Gatherer D, Hilfrich B, Baluchova K, Dargan DJ et al. Sequences of complete human cytomegalovirus genomes from infected cell cultures and clinical specimens. J Gen Virol 2010; 91:605–615 [View Article] [PubMed]
     [Google Scholar]
  40. Jung GS, Kim YY, Kim JI, Ji GY, Jeon JS et al. Full genome sequencing and analysis of human cytomegalovirus strain JHC isolated from a Korean patient. Virus Res 2011; 156:113–120 [View Article] [PubMed]
     [Google Scholar]
  41. Dolan A, Cunningham C, Hector RD, Hassan-Walker AF, Lee L et al. Genetic content of wild-type human cytomegalovirus. J Gen Virol 2004; 85:1301–1312 [View Article] [PubMed]
     [Google Scholar]
  42. Murrell I, Tomasec P, Wilkie GS, Dargan DJ, Davison AJ et al. Impact of sequence variation in the UL128 locus on production of human cytomegalovirus in fibroblast and epithelial cells. J Virol 2013; 87:10489–10500 [View Article] [PubMed]
     [Google Scholar]
  43. Scherer M, Klingl S, Sevvana M, Otto V, Schilling E-M et al. Crystal structure of cytomegalovirus IE1 protein reveals targeting of TRIM family member PML via coiled-coil interactions. PLoS Pathog 2014; 10:e1004512 [View Article] [PubMed]
     [Google Scholar]
  44. Lin Y-T, Chiweshe S, McCormick D, Raper A, Wickenhagen A et al. Human cytomegalovirus evades ZAP detection by suppressing CpG dinucleotides in the major immediate early 1 gene. PLoS Pathog 2020; 16:e1008844 [View Article] [PubMed]
     [Google Scholar]
/content/journal/acmi/10.1099/acmi.0.001104.v3
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Human cytomegalovirus strain-specific differences in protein expression of type I IFN pathway proteins do not impact virus replication
Access Microbiology 8, 001104.v3 (2026); https://doi.org/10.1099/acmi.0.001104.v3
/content/journal/acmi/10.1099/acmi.0.001104.v3
/content/journal/acmi/10.1099/acmi.0.001104.v3
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