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Abstract

To identify new compounds with anti-human cytomegalovirus (HCMV) activity and new anti-HCMV targets, we developed a high-throughput strategy to screen a GlaxoSmithKline Published Kinase Inhibitor Set. This collection contains a range of extensively characterized compounds grouped into chemical families (chemotypes). From our screen, we identified compounds within chemotypes that impede HCMV protein production and identified kinase proteins associated with inhibition of HCMV protein production that are potential novel anti-HCMV targets. We focused our study on a top ‘hit’ in our screen, SB-734117, which we found inhibits productive replication of several HCMV strains. Kinase selectivity data indicated that SB-734117 exhibited polypharmacology and was an inhibitor of several proteins from the AGC and CMCG kinase groups. Using Western blotting, we found that SB-734711 inhibited accumulation of HCMV immediate-early proteins, phosphorylation of cellular proteins involved in immediate-early protein production (cAMP response element-binding protein and histone H3) and histone H3 lysine 36 trimethylation (H3K36me3). Therefore, we identified SB-734117 as a novel anti-HCMV compound and found that inhibition of AGC and CMCG kinase proteins during productive HCMV replication was associated with inhibition of viral protein production and prevented post-translational modification of cellular factors associated with viral protein production.

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2017-04-20
2019-10-23
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References

  1. Mocarski ES, Shenk T, Griffiths PD, Pass RF. Cytomegaloviruses. In Knipe DM, Howley PM. (editors) Fields Virology, 6th ed.vol. 2 New York: Lippincott, Williams & Wilkins; 2015; pp. 1960– 2015
    [Google Scholar]
  2. Krause PR, Bialek SR, Boppana SB, Griffiths PD, Laughlin CA et al. Priorities for CMV vaccine development. Vaccine 2013; 32: 4– 10 [CrossRef] [PubMed]
    [Google Scholar]
  3. Coen DM, Schaffer PA. Antiherpesvirus drugs: a promising spectrum of new drugs and drug targets. Nat Rev Drug Discov 2003; 2: 278– 288 [CrossRef] [PubMed]
    [Google Scholar]
  4. Rogers N. A dormant danger: new therapies target a ubiquitous pathogen known as cytomegalovirus. Nat Med 2015; 21: 1104– 1105 [CrossRef] [PubMed]
    [Google Scholar]
  5. Müller S, Chaikuad A, Gray NS, Knapp S. The ins and outs of selective kinase inhibitor development. Nat Chem Biol 2015; 11: 818– 821 [CrossRef] [PubMed]
    [Google Scholar]
  6. Paolini GV, Shapland RH, van Hoorn WP, Mason JS, Hopkins AL. Global mapping of pharmacological space. Nat Biotechnol 2006; 24: 805– 815 [CrossRef] [PubMed]
    [Google Scholar]
  7. Elkins JM, Fedele V, Szklarz M, Abdul Azeez KR, Salah E et al. Comprehensive characterization of the published kinase inhibitor set. Nat Biotechnol 2016; 34: 95– 103 [CrossRef] [PubMed]
    [Google Scholar]
  8. Drewry DH, Willson TM, Zuercher WJ. Seeding collaborations to advance kinase science with the GSK published kinase inhibitor set (PKIS). Curr Top Med Chem 2014; 14: 340– 342 [CrossRef] [PubMed]
    [Google Scholar]
  9. Polachek WS, Moshrif HF, Franti M, Coen DM, Sreenu VB et al. High-throughput small interfering RNA screening identifies phosphatidylinositol 3-kinase class II alpha as important for production of human cytomegalovirus virions. J Virol 2016; 90: 8360– 8371 [CrossRef] [PubMed]
    [Google Scholar]
  10. Compton T, Nowlin DM, Cooper NR. Initiation of human cytomegalovirus infection requires initial interaction with cell surface heparan sulfate. Virology 1993; 193: 834– 841 [CrossRef] [PubMed]
    [Google Scholar]
  11. Birmingham A, Selfors LM, Forster T, Wrobel D, Kennedy CJ et al. Statistical methods for analysis of high-throughput RNA interference screens. Nat Methods 2009; 6: 569– 575 [CrossRef] [PubMed]
    [Google Scholar]
  12. Zhang JH, Chung TD, Oldenburg KR. A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 1999; 4: 67– 73 [CrossRef] [PubMed]
    [Google Scholar]
  13. Wilkinson GW, Davison AJ, Tomasec P, Fielding CA, Aicheler R et al. Human Cytomegalovirus: taking the strain. Med Microbiol Immunol 2015; 204: 273– 284 [CrossRef] [PubMed]
    [Google Scholar]
  14. White EA, Del Rosario CJ, Sanders RL, Spector DH. The IE2 60-kilodalton and 40-kilodalton proteins are dispensable for human Cytomegalovirus replication but are required for efficient delayed early and late gene expression and production of infectious virus. J Virol 2007; 81: 2573– 2583 [CrossRef] [PubMed]
    [Google Scholar]
  15. Bamford MJ, Alberti MJ, Bailey N, Davies S, Dean DK et al. (1H-imidazo[4,5-c]pyridin-2-yl)-1,2,5-oxadiazol-3-ylamine derivatives: a novel class of potent MSK-1-inhibitors. Bioorg Med Chem Lett 2005; 15: 3402– 3406 [CrossRef] [PubMed]
    [Google Scholar]
  16. Naqvi S, Macdonald A, Mccoy CE, Darragh J, Reith AD et al. Characterization of the cellular action of the MSK inhibitor SB-747651A. Biochem J 2012; 441: 347– 357 [CrossRef] [PubMed]
    [Google Scholar]
  17. Ho CM, Donovan-Banfield IZ, Tan L, Zhang T, Gray NS et al. Inhibition of ikkα by BAY61-3606 reveals IKKα-dependent histone H3 phosphorylation in human cytomegalovirus infected cells. PLoS One 2016; 11: e0150339 [CrossRef] [PubMed]
    [Google Scholar]
  18. Chia YL, Ng CH, Lashmit P, Chu KL, Lew QJ et al. Inhibition of human cytomegalovirus replication by overexpression of CREB1. Antiviral Res 2014; 102: 11– 22 [CrossRef] [PubMed]
    [Google Scholar]
  19. Stinski MF, Thomsen DR, Stenberg RM, Goldstein LC. Organization and expression of the immediate early genes of human cytomegalovirus. J Virol 1983; 46: 1– 14 [PubMed]
    [Google Scholar]
  20. Rodems SM, Clark CL, Spector DH. Separate DNA elements containing ATF/CREB and IE86 binding sites differentially regulate the human cytomegalovirus UL112-113 promoter at early and late times in the infection. J Virol 1998; 72: 2697– 2707 [PubMed]
    [Google Scholar]
  21. 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 [CrossRef] [PubMed]
    [Google Scholar]
  22. Arthur JS. MSK activation and physiological roles. Front Biosci 2008; 13: 5866– 5879 [CrossRef] [PubMed]
    [Google Scholar]
  23. Macdonald N, Welburn JP, Noble ME, Nguyen A, Yaffe MB et al. Molecular basis for the recognition of phosphorylated and phosphoacetylated histone h3 by 14-3-3. Mol Cell 2005; 20: 199– 211 [CrossRef] [PubMed]
    [Google Scholar]
  24. Yamamoto Y, Verma UN, Prajapati S, Kwak YT, Gaynor RB. Histone H3 phosphorylation by IKK-alpha is critical for cytokine-induced gene expression. Nature 2003; 423: 655– 659 [CrossRef] [PubMed]
    [Google Scholar]
  25. Anest V, Hanson JL, Cogswell PC, Steinbrecher KA, Strahl BD et al. A nucleosomal function for IkappaB kinase-alpha in NF-kappaB-dependent gene expression. Nature 2003; 423: 659– 663 [CrossRef] [PubMed]
    [Google Scholar]
  26. Vicent GP, Ballaré C, Nacht AS, Clausell J, Subtil-Rodríguez A et al. Induction of progesterone target genes requires activation of ERK and MSK kinases and phosphorylation of histone H3. Mol Cell 2006; 24: 367– 381 [CrossRef] [PubMed]
    [Google Scholar]
  27. Lo WS, Trievel RC, Rojas JR, Duggan L, Hsu JY et al. Phosphorylation of serine 10 in histone H3 is functionally linked in vitro and in vivo to GCN5-mediated acetylation at lysine 14. Mol Cell 2000; 5: 917– 926 [PubMed] [Crossref]
    [Google Scholar]
  28. 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 [CrossRef] [PubMed]
    [Google Scholar]
  29. Gräff J, Woldemichael BT, Berchtold D, Dewarrat G, Mansuy IM. Dynamic histone marks in the hippocampus and cortex facilitate memory consolidation. Nat Commun 2012; 3: 991 [CrossRef] [PubMed]
    [Google Scholar]
  30. Karam CS, Kellner WA, Takenaka N, Clemmons AW, Corces VG. 14-3-3 mediates histone cross-talk during transcription elongation in Drosophila. PLoS Genet 2010; 6: e1000975 [CrossRef] [PubMed]
    [Google Scholar]
  31. Dong W, Li Y, Gao M, Hu M, Li X et al. IKKα contributes to UVB-induced VEGF expression by regulating AP-1 transactivation. Nucleic Acids Res 2012; 40: 2940– 2955 [CrossRef] [PubMed]
    [Google Scholar]
  32. Lubin FD, Sweatt JD. The IκB kinase regulates chromatin structure during reconsolidation of conditioned fear memories. Neuron 2007; 55: 942– 957 [CrossRef] [PubMed]
    [Google Scholar]
  33. Thorne JL, Ouboussad L, Lefevre PF. Heterochromatin protein 1 gamma and IκB kinase alpha interdependence during tumour necrosis factor gene transcription elongation in activated macrophages. Nucleic Acids Res 2012; 40: 7676– 7689 [CrossRef] [PubMed]
    [Google Scholar]
  34. Meier JL, Keller MJ, Mccoy JJ. Requirement of multiple cis-acting elements in the human cytomegalovirus major immediate-early distal enhancer for viral gene expression and replication. J Virol 2002; 76: 313– 326 [CrossRef] [PubMed]
    [Google Scholar]
  35. Cliffe AR, Arbuckle JH, Vogel JL, Geden MJ, Rothbart SB et al. Neuronal stress pathway mediating a histone methyl/phospho switch is required for herpes simplex virus reactivation. Cell Host Microbe 2015; 18: 649– 658 [CrossRef] [PubMed]
    [Google Scholar]
  36. Stanton RJ, Baluchova K, Dargan DJ, Cunningham C, Sheehy O et al. Reconstruction of the complete human cytomegalovirus genome in a BAC reveals RL13 to be a potent inhibitor of replication. J Clin Invest 2010; 120: 3191– 3208 [CrossRef] [PubMed]
    [Google Scholar]
  37. Loregian A, Coen DM. Selective anti-cytomegalovirus compounds discovered by screening for inhibitors of subunit interactions of the viral polymerase. Chem Biol 2006; 13: 191– 200 [CrossRef] [PubMed]
    [Google Scholar]
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