1887

Journal of General Virology header logo

Volume 104, Issue 9

Repression of the major immediate early promoter of human cytomegalovirus allows transcription from an alternate promoter Open Access

Abstract

Following infection, the human cytomegalovirus (HCMV) genome becomes rapidly associated with host histones which can contribute to the regulation of viral gene expression. This can be seen clearly during HCMV latency where silencing of the major immediate early promoter (MIEP), normally responsible for expression of the key lytic proteins IE72 and IE86, is mediated by histone methylation and recruitment of heterochromatin protein 1. Crucially, reversal of these histone modifications coupled with histone acetylation drives viral reactivation which can be blocked with specific histone acetyltransferase inhibitors (HATi). In lytic infection, a role for HATi is less clear despite the well-established enhancement of viral replication observed with histone deacetylase inhibitors. Here we report that a number of different broad-acting HATi have a minor impact on viral infection and replication during lytic infection with the more overt phenotypes observed at lower multiplicities of infection. However, specific analyses of the regulation of major immediate early (MIE) gene expression reveal that the HATi C646, which targets p300/CBP, transiently repressed MIE gene expression via inhibition of the MIEP but by 24 h post-infection MIE gene expression was rescued due to compensatory activation of an alternative IE promoter, ip2. This suggested that silencing of the MIEP promoted alternative ip2 promoter activity in lytic infection and, consistent with this, ip2 transcription is impaired in cells infected with a recombinant HCMV that does not auto-repress the MIEP at late times of infection. Furthermore, inhibition of the histone methyltransferases known to be responsible for auto-repression is similarly inhibitory to ip2 transcription in wild-type infected cells. We also observe that these discrete transcriptional activities of the MIEP and ip2 promoter are also reflected in reactivation; essentially in cells where the MIEP is silenced, ip2 activity is easier to detect at very early times post-reactivation whereas in cells where robust activation of the MIEP is observed ip2 transcription is reduced or delayed. Finally, we observe that inhibition of pathways demonstrated to be important for reactivation of HCMV in dendritic cells, e.g. in response to IL-6, are preferentially important for activation of the MIEP and not the ip2 promoter. Together, these data add to the hypothesis that the existence of multiple promoters within the MIE region of HCMV can drive reactivation in a cell type- and ligand-specific manner and also suggest that inter-dependent regulatory activity between the two promoters exists.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): cell signalling , chromatin , cytomegalovirus , epigenetics , gene expression and latency
Funding
This study was supported by the:
  • Wellcome Trust (Award WT/204870/Z/16/Z)
    • Principle Award Recipient: MatthewReeves
  • Medical Research Council (Award MR/S00081X/1/MRC)
    • Principle Award Recipient: JohnSinclair
  • Medical Research Council (Award MR/R021384/1)
    • Principle Award Recipient: MatthewReeves
  • Royal Free Charity (Award PhD Studentship)
    • Principle Award Recipient: MatthewReeves
© 2023 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001894
2023-09-13
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/104/9/jgv001894.html?itemId=/content/journal/jgv/10.1099/jgv.0.001894&mimeType=html&fmt=ahah

References

  1. Griffiths P, Reeves M. Pathogenesis of human cytomegalovirus in the immunocompromised host. Nat Rev Microbiol 2021; 19:759–773 [View Article] [PubMed]
     [Google Scholar]
  2. Dupont L, Reeves MB. Cytomegalovirus latency and reactivation: recent insights into an age old problem. Rev Med Virol 2016; 26:75–89 [View Article] [PubMed]
     [Google Scholar]
  3. Limaye AP, Boeckh M. CMV in critically ill patients: pathogen or bystander?. Rev Med Virol 2010; 20:372–379 [View Article] [PubMed]
     [Google Scholar]
  4. Legendre C, Pascual M. Improving outcomes for solid-organ transplant recipients at risk from cytomegalovirus infection: late-onset disease and indirect consequences. Clin Infect Dis 2008; 46:732–740 [View Article] [PubMed]
     [Google Scholar]
  5. Meier JL, Stinski MF. Regulation of human cytomegalovirus immediate-early gene expression. Intervirology 1996; 39:331–342 [View Article] [PubMed]
     [Google Scholar]
  6. Sinclair J. Chromatin structure regulates human cytomegalovirus gene expression during latency, reactivation and lytic infection. Biochim Biophys Acta 2010; 1799:286–295 [View Article] [PubMed]
     [Google Scholar]
  7. Reeves MB, MacAry PA, Lehner PJ, Sissons JGP, Sinclair JH. Latency, chromatin remodeling, and reactivation of human cytomegalovirus in the dendritic cells of healthy carriers. Proc Natl Acad Sci U S A 2005; 102:4140–4145 [View Article] [PubMed]
     [Google Scholar]
  8. Reeves MB, Lehner PJ, Sissons JGP, Sinclair JH. An in vitro model for the regulation of human cytomegalovirus latency and reactivation in dendritic cells by chromatin remodelling. J Gen Virol 2005; 86:2949–2954 [View Article] [PubMed]
     [Google Scholar]
  9. Krishna BA, Miller WE, O’Connor CM. US28: HCMV’s Swiss Army Knife. Viruses 2018; 10:445 [View Article] [PubMed]
     [Google Scholar]
  10. Krishna BA, Poole EL, Jackson SE, Smit MJ, Wills MR et al. Latency-associated expression of human cytomegalovirus US28 attenuates cell signaling pathways to maintain latent infection. mBio 2017; 8:e01754-17 [View Article] [PubMed]
     [Google Scholar]
  11. Hahn G, Jores R, Mocarski ES. Cytomegalovirus remains latent in a common precursor of dendritic and myeloid cells. Proc Natl Acad Sci 1998; 95:3937–3942 [View Article] [PubMed]
     [Google Scholar]
  12. Taylor-Wiedeman J, Sissons P, Sinclair J. Induction of endogenous human cytomegalovirus gene expression after differentiation of monocytes from healthy carriers. J Virol 1994; 68:1597–1604 [View Article] [PubMed]
     [Google Scholar]
  13. Reeves MB, Compton T. Inhibition of inflammatory interleukin-6 activity via extracellular signal-regulated kinase-mitogen-activated protein kinase signaling antagonizes human cytomegalovirus reactivation from dendritic cells. J Virol 2011; 85:12750–12758 [View Article] [PubMed]
     [Google Scholar]
  14. Hargett D, Shenk TE. Experimental human cytomegalovirus latency in CD14+ monocytes. Proc Natl Acad Sci U S A 2010; 107:20039–20044 [View Article] [PubMed]
     [Google Scholar]
  15. Prösch S, Wuttke R, Krüger DH, Volk H-D. NF-kappaB--a potential therapeutic target for inhibition of human cytomegalovirus (re)activation?. Biol Chem 2002; 383:1601–1609 [View Article] [PubMed]
     [Google Scholar]
  16. Kew VG, Yuan J, Meier J, Reeves MB. Mitogen and stress activated kinases act co-operatively with CREB during the induction of human cytomegalovirus immediate-early gene expression from latency. PLoS Pathog 2014; 10:e1004195 [View Article] [PubMed]
     [Google Scholar]
  17. Reeves MB. Cell signaling and cytomegalovirus reactivation: what do Src family kinases have to do with it?. Biochem Soc Trans 2020; 48:667–675 [View Article] [PubMed]
     [Google Scholar]
  18. Dupont L, Du L, Poulter M, Choi S, McIntosh M et al. Src family kinase activity drives cytomegalovirus reactivation by recruiting MOZ histone acetyltransferase activity to the viral promoter. J Biol Chem 2019; 294:12901–12910 [View Article] [PubMed]
     [Google Scholar]
  19. Krishna BA, Lau B, Jackson SE, Wills MR, Sinclair JH et al. Transient activation of human cytomegalovirus lytic gene expression during latency allows cytotoxic T cell killing of latently infected cells. Sci Rep 2016; 6:24674 [View Article] [PubMed]
     [Google Scholar]
  20. Groves IJ, Jackson SE, Poole EL, Nachshon A, Rozman B et al. Bromodomain proteins regulate human cytomegalovirus latency and reactivation allowing epigenetic therapeutic intervention. Proc Natl Acad Sci U S A 2021; 118:e2023025118 [View Article] [PubMed]
     [Google Scholar]
  21. Arend KC, Ziehr B, Vincent HA, Moorman NJ. Multiple transcripts encode full-length human Cytomegalovirus IE1 and IE2 proteins during lytic infection. J Virol 2016; 90:8855–8865 [View Article] [PubMed]
     [Google Scholar]
  22. Collins-McMillen D, Rak M, Buehler JC, Igarashi-Hayes S, Kamil JP et al. Alternative promoters drive human cytomegalovirus reactivation from latency. Proc Natl Acad Sci U S A 2019; 116:17492–17497 [View Article] [PubMed]
     [Google Scholar]
  23. Hale AE, Collins-McMillen D, Lenarcic EM, Igarashi S, Kamil JP et al. FOXO transcription factors activate alternative major immediate early promoters to induce human cytomegalovirus reactivation. Proc Natl Acad Sci U S A 2020; 117:18764–18770 [View Article] [PubMed]
     [Google Scholar]
  24. Mason R, Groves IJ, Wills MR, Sinclair JH, Reeves MB. Human cytomegalovirus major immediate early transcripts arise predominantly from the canonical major immediate early promoter in reactivating progenitor-derived dendritic cells. J Gen Virol 2020; 101:635–644 [View Article] [PubMed]
     [Google Scholar]
  25. Collins-McMillen D, Kamil J, Moorman N, Goodrum F. Control of immediate early gene expression for human Cytomegalovirus reactivation. Front Cell Infect Microbiol 2020; 10:476 [View Article] [PubMed]
     [Google Scholar]
  26. Albright ER, Walter RM, Saffert RT, Kalejta RF. NFκB and cyclic AMP response element sites mediate the valproic acid and UL138 responsiveness of the human Cytomegalovirus major immediate early enhancer and promoter. J Virol 2023; 97:e0002923 [View Article] [PubMed]
     [Google Scholar]
  27. Keller MJ, Wu AW, Andrews JI, McGonagill PW, Tibesar EE et al. Reversal of human cytomegalovirus major immediate-early enhancer/promoter silencing in quiescently infected cells via the cyclic AMP signaling pathway. J Virol 2007; 81:6669–6681 [View Article] [PubMed]
     [Google Scholar]
  28. Keller MJ, Wheeler DG, Cooper E, Meier JL. Role of the human cytomegalovirus major immediate-early promoter’s 19-base-pair-repeat cyclic AMP-response element in acutely infected cells. J Virol 2003; 77:6666–6675 [View Article] [PubMed]
     [Google Scholar]
  29. Krishna BA, Wass AB, Dooley AL, O’Connor CM. CMV-encoded GPCR pUL33 activates CREB and facilitates its recruitment to the MIE locus for efficient viral reactivation. J Cell Sci 2021; 134:jcs254268 [View Article] [PubMed]
     [Google Scholar]
  30. Reeves MB. Chromatin-mediated regulation of cytomegalovirus gene expression. Virus Res 2011; 157:134–143 [View Article] [PubMed]
     [Google Scholar]
  31. Cuevas-Bennett C, Shenk T. Dynamic histone H3 acetylation and methylation at human cytomegalovirus promoters during replication in fibroblasts. J Virol 2008; 82:9525–9536 [View Article] [PubMed]
     [Google Scholar]
  32. Nitzsche A, Paulus C, Nevels M. Temporal dynamics of cytomegalovirus chromatin assembly in productively infected human cells. J Virol 2008; 82:11167–11180 [View Article] [PubMed]
     [Google Scholar]
  33. Groves IJ, Reeves MB, Sinclair JH. Lytic infection of permissive cells with human cytomegalovirus is regulated by an intrinsic “pre-immediate-early” repression of viral gene expression mediated by histone post-translational modification. J Gen Virol 2009; 90:2364–2374 [View Article] [PubMed]
     [Google Scholar]
  34. Michaelis M, Suhan T, Reinisch A, Reisenauer A, Fleckenstein C et al. Increased replication of human cytomegalovirus in retinal pigment epithelial cells by valproic acid depends on histone deacetylase inhibition. Invest Ophthalmol Vis Sci 2005; 46:3451–3457 [View Article] [PubMed]
     [Google Scholar]
  35. Woodhall DL, Groves IJ, Reeves MB, Wilkinson G, Sinclair JH. Human Daxx-mediated repression of human cytomegalovirus gene expression correlates with a repressive chromatin structure around the major immediate early promoter. J Biol Chem 2006; 281:37652–37660 [View Article] [PubMed]
     [Google Scholar]
  36. Nevels M, Paulus C, Shenk T. Human cytomegalovirus immediate-early 1 protein facilitates viral replication by antagonizing histone deacetylation. Proc Natl Acad Sci U S A 2004; 101:17234–17239 [View Article] [PubMed]
     [Google Scholar]
  37. Mocarski ES, Kemble GW, Lyle JM, Greaves RF. A deletion mutant in the human cytomegalovirus gene encoding IE1(491aa) is replication defective due to A failure in autoregulation. Proc Natl Acad Sci U S A 1996; 93:11321–11326 [View Article] [PubMed]
     [Google Scholar]
  38. Vogel JL, Kristie TM. The dynamics of HCF-1 modulation of herpes simplex virus chromatin during initiation of infection. Viruses 2013; 5:1272–1291 [View Article] [PubMed]
     [Google Scholar]
  39. Gan X, Wang H, Yu Y, Yi W, Zhu S et al. Epigenetically repressing human cytomegalovirus lytic infection and reactivation from latency in THP-1 model by targeting H3K9 and H3K27 histone demethylases. PLoS One 2017; 12:e0175390 [View Article] [PubMed]
     [Google Scholar]
  40. Liang Y, Vogel JL, Narayanan A, Peng H, Kristie TM. Inhibition of the histone demethylase LSD1 blocks alpha-herpesvirus lytic replication and reactivation from latency. Nat Med 2009; 15:1312–1317 [View Article] [PubMed]
     [Google Scholar]
  41. Reeves M, Murphy J, Greaves R, Fairley J, Brehm A et al. Autorepression of the human cytomegalovirus major immediate-early promoter/enhancer at late times of infection is mediated by the recruitment of chromatin remodeling enzymes by IE86. J Virol 2006; 80:9998–10009 [View Article] [PubMed]
     [Google Scholar]
  42. Cherrington JM, Khoury EL, Mocarski ES. Human cytomegalovirus ie2 negatively regulates alpha gene expression via a short target sequence near the transcription start site. J Virol 1991; 65:887–896 [View Article] [PubMed]
     [Google Scholar]
  43. Lee KK, Workman JL. Histone acetyltransferase complexes: one size doesn’t fit all. Nat Rev Mol Cell Biol 2007; 8:284–295 [View Article] [PubMed]
     [Google Scholar]
  44. Elmallah MIY, Micheau O. Epigenetic Regulation of TRAIL Signaling: Implication for Cancer Therapy. Cancers 2019; 11:850 [View Article] [PubMed]
     [Google Scholar]
  45. Li R, Zhu J, Xie Z, Liao G, Liu J et al. Conserved herpesvirus kinases target the DNA damage response pathway and TIP60 histone acetyltransferase to promote virus replication. Cell Host Microbe 2011; 10:390–400 [View Article] [PubMed]
     [Google Scholar]
  46. Bryant LA, Mixon P, Davidson M, Bannister AJ, Kouzarides T et al. The human cytomegalovirus 86-kilodalton major immediate-early protein interacts physically and functionally with histone acetyltransferase P/CAF. J Virol 2000; 74:7230–7237 [View Article] [PubMed]
     [Google Scholar]
  47. Chimenti F, Bizzarri B, Maccioni E, Secci D, Bolasco A et al. A novel histone acetyltransferase inhibitor modulating Gcn5 network: cyclopentylidene-[4-(4’-chlorophenyl)thiazol-2-yl)hydrazone. J Med Chem 2009; 52:530–536 [View Article] [PubMed]
     [Google Scholar]
  48. Perera MR, Wills MR, Sinclair JH. HCMV antivirals and strategies to target the latent reservoir. Viruses 2021; 13:817 [View Article] [PubMed]
     [Google Scholar]
  49. Spector BM, Parida M, Li M, Ball CB, Meier JL et al. Differences in RNA polymerase II complexes and their interactions with surrounding chromatin on human and cytomegalovirus genomes. Nat Commun 2022; 13:2006 [View Article] [PubMed]
     [Google Scholar]
  50. Tarrant-Elorza M, Rossetto CC, Pari GS. Maintenance and replication of the human cytomegalovirus genome during latency. Cell Host Microbe 2014; 16:43–54 [View Article] [PubMed]
     [Google Scholar]
  51. Albright ER, Kalejta RF. Canonical and variant forms of histone H3 are deposited onto the human cytomegalovirus genome during lytic and latent infections. J Virol 2016; 90:10309–10320 [View Article] [PubMed]
     [Google Scholar]
  52. Rauwel B, Jang SM, Cassano M, Kapopoulou A, Barde I et al. Release of human cytomegalovirus from latency by a KAP1/TRIM28 phosphorylation switch. eLife 2015; 4:e06068 [View Article] [PubMed]
     [Google Scholar]
  53. Krishna BA, Wass AB, O’Connor CM. Activator protein-1 transactivation of the major immediate early locus is a determinant of cytomegalovirus reactivation from latency. Proc Natl Acad Sci U S A 2020; 117:20860–20867 [View Article] [PubMed]
     [Google Scholar]
  54. Buehler J, Carpenter E, Zeltzer S, Igarashi S, Rak M et al. Host signaling and EGR1 transcriptional control of human cytomegalovirus replication and latency. PLoS Pathog 2019; 15:e1008037 [View Article] [PubMed]
     [Google Scholar]
  55. Buehler J, Zeltzer S, Reitsma J, Petrucelli A, Umashankar M et al. Opposing regulation of the EGF receptor: a molecular switch controlling cytomegalovirus latency and replication. PLoS Pathog 2016; 12:e1005655 [View Article] [PubMed]
     [Google Scholar]
  56. Akasaki Y, Alvarez-Garcia O, Saito M, Caramés B, Iwamoto Y et al. FoxO transcription factors support oxidative stress resistance in human chondrocytes. Arthritis Rheumatol 2014; 66:3349–3358 [View Article] [PubMed]
     [Google Scholar]
  57. Sengupta A, Molkentin JD, Paik J-H, DePinho RA, Yutzey KE. FoxO transcription factors promote cardiomyocyte survival upon induction of oxidative stress. J Biol Chem 2011; 286:7468–7478 [View Article] [PubMed]
     [Google Scholar]
  58. Gille H, Strahl T, Shaw PE. Activation of ternary complex factor Elk-1 by stress-activated protein kinases. Curr Biol 1995; 5:1191–1200 [View Article] [PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001894
Loading
Repression of the major immediate early promoter of human cytomegalovirus allows transcription from an alternate promoter
J. Gen. Virol. 104, 001894 (2023); https://doi.org/10.1099/jgv.0.001894
/content/journal/jgv/10.1099/jgv.0.001894
/content/journal/jgv/10.1099/jgv.0.001894
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error