1887

Journal of General Virology header logo

Volume 90, Issue 8

Cellular kinase inhibitors that suppress enterovirus replication have a conserved target in viral protein 3A similar to that of enviroxime Free

Abstract

Previously, we identified a cellular kinase inhibitor, GW5074, that inhibits poliovirus (PV) and enterovirus 71 replication strongly, although its target has remained unknown. To identify the target of GW5074, we searched for cellular kinase inhibitors that have anti-enterovirus activity similar or related to that of GW5074. With this aim, we performed screenings to identify cellular kinase inhibitors that could inhibit PV replication cooperatively with GW5074 or synthetically in the absence of GW5074. We identified MEK1/2 inhibitors (SL327 and U0126), an EGFR inhibitor (AG1478) and a phosphatidylinositol 3-kinase inhibitor (wortmannin) as compounds with a cooperative inhibitory effect with GW5074, and an Akt1/2 inhibitor (Akt inhibitor VIII) as a compound with a synthetic inhibitory effect with MEK1/2 inhibitors and AG1478. Individual treatment with the identified kinase inhibitors did not affect PV replication significantly, but combined treatment with MEK1/2 inhibitor, AG1478 and Akt1/2 inhibitor suppressed the replication synthetically. The effect of AG1478 in this synthetic inhibition was compensated by other receptor tyrosine kinase inhibitors (IGF-1R inhibitor II and Flt3 inhibitor II). We isolated mutants resistant to Flt3 inhibitor II and GW5074 and found that these mutants had cross-resistance to each treatment. These mutants had a common mutation in viral protein 3A that results in an amino acid change at position 70 (Ala to Thr), a mutation that was previously identified in mutants resistant to a potent anti-enterovirus compound, enviroxime. These results suggest that cellular kinase inhibitors and enviroxime have a conserved target in viral protein 3A to suppress enterovirus replication.

  • Received:
  • Accepted:
  • Published Online:
SGM
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.012096-0
2009-08-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/8/1869.html?itemId=/content/journal/jgv/10.1099/vir.0.012096-0&mimeType=html&fmt=ahah

References

  1. Arita M., Shimizu H., Nagata N., Ami Y., Suzaki Y., Sata T., Iwasaki T., Miyamura T. 2005a; Temperature-sensitive mutants of enterovirus 71 show attenuation in cynomolgus monkeys. J Gen Virol 86:1391–1401 [CrossRef]
     [Google Scholar]
  2. Arita M., Zhu S. L., Yoshida H., Yoneyama T., Miyamura T., Shimizu H. 2005b; A Sabin 3-derived poliovirus recombinant contained a sequence homologous with indigenous human enterovirus species C in the viral polymerase coding region. J Virol 79:12650–12657 [CrossRef]
     [Google Scholar]
  3. Arita M., Nagata N., Sata T., Miyamura T., Shimizu H. 2006; Quantitative analysis of poliomyelitis-like paralysis in mice induced by a poliovirus replicon. J Gen Virol 87:3317–3327 [CrossRef]
     [Google Scholar]
  4. Arita M., Nagata N., Iwata N., Ami Y., Suzaki Y., Mizuta K., Iwasaki T., Sata T., Wakita T., Shimizu H. 2007; An attenuated strain of enterovirus 71 belonging to genotype A showed a broad spectrum of antigenicity with attenuated neurovirulence in cynomolgus monkeys. J Virol 81:9386–9395 [CrossRef]
     [Google Scholar]
  5. Arita M., Ami Y., Wakita T., Shimizu H. 2008a; Cooperative effect of the attenuation determinants derived from poliovirus Sabin 1 strain is essential for attenuation of enterovirus 71 in the NOD/SCID mouse infection model. J Virol 82:1787–1797 [CrossRef]
     [Google Scholar]
  6. Arita M., Wakita T., Shimizu H. 2008b; Characterization of pharmacologically active compounds that inhibit poliovirus and enterovirus 71 infectivity. J Gen Virol 89:2518–2530 [CrossRef]
     [Google Scholar]
  7. Autret A., Martin-Latil S., Brisac C., Mousson L., Colbere-Garapin F., Blondel B. 2008; Early phosphatidylinositol 3-kinase/Akt pathway activation limits poliovirus-induced JNK-mediated cell death. J Virol 82:3796–3802 [CrossRef]
     [Google Scholar]
  8. Bain J., Plater L., Elliott M., Shpiro N., Hastie J., McLauchlan H., Klevernic I., Arthur S., Alessi D., Cohen P. 2007; The selectivity of protein kinase inhibitors; a further update. Biochem J 408:297–315 [CrossRef]
     [Google Scholar]
  9. Belov G. A., Feng Q., Nikovics K., Jackson C. L., Ehrenfeld E. 2008; A critical role of a cellular membrane traffic protein in poliovirus RNA replication. PLoS Pathog 4:e1000216 [CrossRef]
     [Google Scholar]
  10. Bodian D. 1949; Histopathologic basis of clinical findings in poliomyelitis. Am J Med 6:563–578 [CrossRef]
     [Google Scholar]
  11. Bordeleau M. E., Mori A., Oberer M., Lindqvist L., Chard L. S., Higa T., Belsham G. J., Wagner G., Tanaka J., Pelletier J. 2006; Functional characterization of IRESes by an inhibitor of the RNA helicase eIF4A. Nat Chem Biol 2:213–220 [CrossRef]
     [Google Scholar]
  12. Brown-Augsburger P., Vance L. M., Malcolm S. K., Hsiung H., Smith D. P., Heinz B. A. 1999; Evidence that enviroxime targets multiple components of the rhinovirus 14 replication complex. Arch Virol 144:1569–1585 [CrossRef]
     [Google Scholar]
  13. Chang L. Y., Lin T. Y., Hsu K. H., Huang Y. C., Lin K. L., Hsueh C., Shih S. R., Ning H. C., Hwang M. S. other authors 1999; Clinical features and risk factors of pulmonary oedema after enterovirus-71-related hand, foot, and mouth disease. Lancet 354:1682–1686 [CrossRef]
     [Google Scholar]
  14. Chen H. F., Chang M. H., Chiang B. L., Jeng S. T. 2006; Oral immunization of mice using transgenic tomato fruit expressing VP1 protein from enterovirus 71. Vaccine 24:2944–2951 [CrossRef]
     [Google Scholar]
  15. Chiu C. H., Chu C., He C. C., Lin T. Y. 2006; Protection of neonatal mice from lethal enterovirus 71 infection by maternal immunization with attenuated Salmonella enterica serovar Typhimurium expressing VP1 of enterovirus 71. Microbes Infect 8:1671–1678 [CrossRef]
     [Google Scholar]
  16. Chumakov M., Voroshilova M., Shindarov L., Lavrova I., Gracheva L., Koroleva G., Vasilenko S., Brodvarova I., Nikolova M. other authors 1979; Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria. Arch Virol 60:329–340 [CrossRef]
     [Google Scholar]
  17. Crotty S., Saleh M. C., Gitlin L., Beske O., Andino R. 2004; The poliovirus replication machinery can escape inhibition by an antiviral drug that targets a host cell protein. J Virol 78:3378–3386 [CrossRef]
     [Google Scholar]
  18. Cuconati A., Molla A., Wimmer E. 1998; Brefeldin A inhibits cell-free, de novo synthesis of poliovirus. J Virol 72:6456–6464
     [Google Scholar]
  19. DeFeo-Jones D., Barnett S. F., Fu S., Hancock P. J., Haskell K. M., Leander K. R., McAvoy E., Robinson R. G., Duggan M. E. other authors 2005; Tumor cell sensitization to apoptotic stimuli by selective inhibition of specific Akt/PKB family members. Mol Cancer Ther 4:271–279
     [Google Scholar]
  20. De Palma A. M., Thibaut H. J., van der Linden L., Lanke K., Heggermont W., Ireland S., Andrews R., Arimilli M., Al-Tel T. other authors 2009; Mutations in the non-structural protein 3A confer resistance to the novel enterovirus replication inhibitor TTP-8307. Antimicrob Agents Chemother 53:1850–1857 [CrossRef]
     [Google Scholar]
  21. Esfandiarei M., Luo H., Yanagawa B., Suarez A., Dabiri D., Zhang J., McManus B. M. 2004; Protein kinase B/Akt regulates coxsackievirus B3 replication through a mechanism which is not caspase dependent. J Virol 78:4289–4298 [CrossRef]
     [Google Scholar]
  22. Favata M. F., Horiuchi K. Y., Manos E. J., Daulerio A. J., Stradley D. A., Feeser W. S., Van Dyk D. E., Pitts W. J., Earl R. A. other authors 1998; Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J Biol Chem 273:18623–18632 [CrossRef]
     [Google Scholar]
  23. Geller R., Vignuzzi M., Andino R., Frydman J. 2007; Evolutionary constraints on chaperone-mediated folding provide an antiviral approach refractory to development of drug resistance. Genes Dev 21:195–205 [CrossRef]
     [Google Scholar]
  24. Heinz B. A., Vance L. M. 1995; The antiviral compound enviroxime targets the 3A coding region of rhinovirus and poliovirus. J Virol 69:4189–4197
     [Google Scholar]
  25. Ho M., Chen E. R., Hsu K. H., Twu S. J., Chen K. T., Tsai S. F., Wang J. R., Shih S. R. 1999; An epidemic of enterovirus 71 infection in Taiwan. N Engl J Med 341:929–935 [CrossRef]
     [Google Scholar]
  26. Huang C. C., Liu C. C., Chang Y. C., Chen C. Y., Wang S. T., Yeh T. F. 1999; Neurologic complications in children with enterovirus 71 infection. N Engl J Med 341:936–942 [CrossRef]
     [Google Scholar]
  27. Irurzun A., Perez L., Carrasco L. 1992; Involvement of membrane traffic in the replication of poliovirus genomes: effects of brefeldin A. Virology 191:166–175 [CrossRef]
     [Google Scholar]
  28. Isaacson M. K., Feire A. L., Compton T. 2007; Epidermal growth factor receptor is not required for human cytomegalovirus entry or signaling. J Virol 81:6241–6247 [CrossRef]
     [Google Scholar]
  29. Komatsu H., Shimizu Y., Takeuchi Y., Ishiko H., Takada H. 1999; Outbreak of severe neurologic involvement associated with enterovirus 71 infection. Pediatr Neurol 20:17–23 [CrossRef]
     [Google Scholar]
  30. Krenn B. M., Holzer B., Gaudernak E., Triendl A., van Kuppeveld F. J., Seipelt J. 2005; Inhibition of polyprotein processing and RNA replication of human rhinovirus by pyrrolidine dithiocarbamate involves metal ions. J Virol 79:13892–13899 [CrossRef]
     [Google Scholar]
  31. Krenn B. M., Gaudernak E., Holzer B., Lanke K., Van Kuppeveld F. J., Seipelt J. 2009; Antiviral activity of zinc ionophores pyrithione and hinokitiol against picornaviral infections. J Virol 83:58–64 [CrossRef]
     [Google Scholar]
  32. Lanke K., Krenn B. M., Melchers W. J., Seipelt J., van Kuppeveld F. J. 2007; PDTC inhibits picornavirus polyprotein processing and RNA replication by transporting zinc ions into cells. J Gen Virol 88:1206–1217 [CrossRef]
     [Google Scholar]
  33. Leleu X., Jia X., Runnels J., Ngo H. T., Moreau A. S., Farag M., Spencer J. A., Pitsillides C. M., Hatjiharissi E. other authors 2007; The Akt pathway regulates survival and homing in Waldenstrom macroglobulinemia. Blood 110:4417–4426 [CrossRef]
     [Google Scholar]
  34. Liu M. L., Lee Y. P., Wang Y. F., Lei H. Y., Liu C. C., Wang S. M., Su I. J., Wang J. R., Yeh T. M. other authors 2005; Type I interferons protect mice against enterovirus 71 infection. J Gen Virol 86:3263–3269 [CrossRef]
     [Google Scholar]
  35. Liu C. C., Lian W. C., Butler M., Wu S. C. 2007; High immunogenic enterovirus 71 strain and its production using serum-free microcarrier Vero cell culture. Vaccine 25:19–24 [CrossRef]
     [Google Scholar]
  36. Logie L., Ruiz-Alcaraz A. J., Keane M., Woods Y. L., Bain J., Marquez R., Alessi D. R., Sutherland C. 2007; Characterization of a protein kinase B inhibitor in vitro and in insulin-treated liver cells. Diabetes 56:2218–2227 [CrossRef]
     [Google Scholar]
  37. Lum L. C., Wong K. T., Lam S. K., Chua K. B., Goh A. Y., Lim W. L., Ong B. B., Paul G., AbuBakar S., Lambert M. 1998; Fatal enterovirus 71 encephalomyelitis. J Pediatr 133:795–798 [CrossRef]
     [Google Scholar]
  38. Luo H., Yanagawa B., Zhang J., Luo Z., Zhang M., Esfandiarei M., Carthy C., Wilson J. E., Yang D., McManus B. M. 2002; Coxsackievirus B3 replication is reduced by inhibition of the extracellular signal-regulated kinase (ERK) signaling pathway. J Virol 76:3365–3373 [CrossRef]
     [Google Scholar]
  39. Macejak D. G., Sarnow P. 1992; Association of heat shock protein 70 with enterovirus capsid precursor P1 in infected human cells. J Virol 66:1520–1527
     [Google Scholar]
  40. Maynell L. A., Kirkegaard K., Klymkowsky M. W. 1992; Inhibition of poliovirus RNA synthesis by brefeldin A. J Virol 66:1985–1994
     [Google Scholar]
  41. McMinn P. C. 2002; An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS Microbiol Rev 26:91–107 [CrossRef]
     [Google Scholar]
  42. Nijhuis M., van Maarseveen N., Schuurman R., Verkuijlen S., de Vos M., Hendriksen K., van Loon A. M. 2002; Rapid and sensitive routine detection of all members of the genus Enterovirus in different clinical specimens by real-time PCR. J Clin Microbiol 40:3666–3670 [CrossRef]
     [Google Scholar]
  43. Nkowane B. M., Wassilak S. G., Orenstein W. A., Bart K. J., Schonberger L. B., Hinman A. R., Kew O. M. 1987; Vaccine-associated paralytic poliomyelitis. United States: 1973 through 1984. JAMA 257:1335–1340 [CrossRef]
     [Google Scholar]
  44. Okada T., Kawano Y., Sakakibara T., Hazeki O., Ui M. 1994; Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes. Studies with a selective inhibitor wortmannin. J Biol Chem 269:3568–3573
     [Google Scholar]
  45. Rusnak D. W., Affleck K., Cockerill S. G., Stubberfield C., Harris R., Page M., Smith K. J., Guntrip S. B., Carter M. C. other authors 2001; The characterization of novel, dual ErbB-2/EGFR, tyrosine kinase inhibitors: potential therapy for cancer. Cancer Res 61:7196–7203
     [Google Scholar]
  46. Sabin A. B. 1965; Oral poliovirus vaccine. History of its development and prospects for eradication of poliomyelitis. JAMA 194:872–876 [CrossRef]
     [Google Scholar]
  47. Salk J. E., Bazeley P. L., Bennett B. L., Krech U., Lewis L. J., Ward E. N., Youngner J. S. 1954; Studies in human subjects on active immunization against poliomyelitis. II. A practical means for inducing and maintaining antibody formation. Am J Public Health Nations Health 44:994–1009 [CrossRef]
     [Google Scholar]
  48. Shih S. R., Tsai M. C., Tseng S. N., Won K. F., Shia K. S., Li W. T., Chern J. H., Chen G. W., Lee C. C. other authors 2004; Mutation in enterovirus 71 capsid protein VP1 confers resistance to the inhibitory effects of pyridyl imidazolidinone. Antimicrob Agents Chemother 48:3523–3529 [CrossRef]
     [Google Scholar]
  49. Si X., McManus B. M., Zhang J., Yuan J., Cheung C., Esfandiarei M., Suarez A., Morgan A., Luo H. 2005; Pyrrolidine dithiocarbamate reduces coxsackievirus B3 replication through inhibition of the ubiquitin–proteasome pathway. J Virol 79:8014–8023 [CrossRef]
     [Google Scholar]
  50. Soeda A., Inagaki A., Oka N., Ikegame Y., Aoki H., Yoshimura S., Nakashima S., Kunisada T., Iwama T. 2008; Epidermal growth factor plays a crucial role in mitogenic regulation of human brain tumor stem cells. J Biol Chem 283:10958–10966 [CrossRef]
     [Google Scholar]
  51. Svitkin Y. V., Hahn H., Gingras A. C., Palmenberg A. C., Sonenberg N. 1998; Rapamycin and wortmannin enhance replication of a defective encephalomyocarditis virus. J Virol 72:5811–5819
     [Google Scholar]
  52. Tan C. S., Cardosa M. J. 2007; High-titred neutralizing antibodies to human enterovirus 71 preferentially bind to the N-terminal portion of the capsid protein VP1. Arch Virol 152:1069–1073 [CrossRef]
     [Google Scholar]
  53. Tung W. S., Bakar S. A., Sekawi Z., Rosli R. 2007; DNA vaccine constructs against enterovirus 71 elicit immune response in mice. Genet Vaccines Ther 5:6 [CrossRef]
     [Google Scholar]
  54. Upla P., Marjomaki V., Nissinen L., Nylund C., Waris M., Hyypia T., Heino J. 2008; Calpain 1 and 2 are required for RNA replication of echovirus 1. J Virol 82:1581–1590 [CrossRef]
     [Google Scholar]
  55. Wang S. M., Liu C. C., Tseng H. W., Wang J. R., Huang C. C., Chen Y. J., Yang Y. J., Lin S. J., Yeh T. F. 1999; Clinical spectrum of enterovirus 71 infection in children in southern Taiwan, with an emphasis on neurological complications. Clin Infect Dis 29:184–190 [CrossRef]
     [Google Scholar]
  56. Wang S. M., Lei H. Y., Huang K. J., Wu J. M., Wang J. R., Yu C. K., Su I. J., Liu C. C. 2003; Pathogenesis of enterovirus 71 brainstem encephalitis in pediatric patients: roles of cytokines and cellular immune activation in patients with pulmonary edema. J Infect Dis 188:564–570 [CrossRef]
     [Google Scholar]
  57. Wikel J. H., Paget C. J., DeLong D. C., Nelson J. D., Wu C. Y., Paschal J. W., Dinner A., Templeton R. J., Chaney M. O. other authors 1980; Synthesis of syn and anti isomers of 6-[[(hydroxyimino)phenyl]methyl]-1-[(1-methylethyl)sulfonyl]-1H-benzimidazol-2-amine. Inhibitors of rhinovirus multiplication. J Med Chem 23:368–372 [CrossRef]
     [Google Scholar]
  58. Wu T. C., Wang Y. F., Lee Y. P., Wang J. R., Liu C. C., Wang S. M., Lei H. Y., Su I. J., Yu C. K. 2007; Immunity to avirulent enterovirus 71 and coxsackie A16 virus protects against enterovirus 71 infection in mice. J Virol 81:10310–10315 [CrossRef]
     [Google Scholar]
  59. Yu C. K., Chen C. C., Chen C. L., Wang J. R., Liu C. C., Yan J. J., Su I. J. 2000; Neutralizing antibody provided protection against enterovirus type 71 lethal challenge in neonatal mice. J Biomed Sci 7:523–528 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.012096-0
Loading
Cellular kinase inhibitors that suppress enterovirus replication have a conserved target in viral protein 3A similar to that of enviroxime
J. Gen. Virol. 90, 1869 (2009); https://doi.org/10.1099/vir.0.012096-0
/content/journal/jgv/10.1099/vir.0.012096-0
/content/journal/jgv/10.1099/vir.0.012096-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

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