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. 1990 Sep;64(9):4573–4577. doi: 10.1128/jvi.64.9.4573-4577.1990

Cleavage of dengue virus NS1-NS2A requires an octapeptide sequence at the C terminus of NS1.

H Hori 1, C J Lai 1
PMCID: PMC247931  PMID: 2143546

Abstract

The length of amino acid sequence at the NS1-NS2A juncture of dengue virus that is required for specific cleavage effected by the cis-acting function of NS2A was identified by deletion analysis. Recombinant DNA sequences of NS1-NS2A, each containing a deletion in NS1 followed by a sequence of 3 to 20 amino acids at the C terminus of NS1 preceding the cleavage site, were constructed and expressed with vaccinia virus as a vector. The NS1 product of recombinant vaccinia virus-infected cells was immunoprecipitated and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The occurrence of cleavage between NS1 and NS2A was indicated by the appearance of shortened NS1. Failure to cleave this site yielded a large NS1-NS2A fusion protein. This analysis indicated that a minimum length of eight amino acids at the NS1 C terminus preceding the NS1-NS2A juncture is required for cleavage to take place. Comparison of this eight-amino-acid sequence of the NS1 C terminus of dengue type 4 virus with the analogous sequences of 12 other flaviviruses suggests that the consensus cleavage site sequence is as follows: (table; see text)

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bell J. R., Kinney R. M., Trent D. W., Lenches E. M., Dalgarno L., Strauss J. H. Amino-terminal amino acid sequences of structural proteins of three flaviviruses. Virology. 1985 May;143(1):224–229. doi: 10.1016/0042-6822(85)90110-2. [DOI] [PubMed] [Google Scholar]
  2. Biedrzycka A., Cauchi M. R., Bartholomeusz A., Gorman J. J., Wright P. J. Characterization of protease cleavage sites involved in the formation of the envelope glycoprotein and three non-structural proteins of dengue virus type 2, New Guinea C strain. J Gen Virol. 1987 May;68(Pt 5):1317–1326. doi: 10.1099/0022-1317-68-5-1317. [DOI] [PubMed] [Google Scholar]
  3. Carrington J. C., Cary S. M., Dougherty W. G. Mutational analysis of tobacco etch virus polyprotein processing: cis and trans proteolytic activities of polyproteins containing the 49-kilodalton proteinase. J Virol. 1988 Jul;62(7):2313–2320. doi: 10.1128/jvi.62.7.2313-2320.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Castle E., Leidner U., Nowak T., Wengler G., Wengler G. Primary structure of the West Nile flavivirus genome region coding for all nonstructural proteins. Virology. 1986 Feb;149(1):10–26. doi: 10.1016/0042-6822(86)90082-6. [DOI] [PubMed] [Google Scholar]
  5. Castle E., Nowak T., Leidner U., Wengler G., Wengler G. Sequence analysis of the viral core protein and the membrane-associated proteins V1 and NV2 of the flavivirus West Nile virus and of the genome sequence for these proteins. Virology. 1985 Sep;145(2):227–236. doi: 10.1016/0042-6822(85)90156-4. [DOI] [PubMed] [Google Scholar]
  6. Chakrabarti S., Brechling K., Moss B. Vaccinia virus expression vector: coexpression of beta-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol. 1985 Dec;5(12):3403–3409. doi: 10.1128/mcb.5.12.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coia G., Parker M. D., Speight G., Byrne M. E., Westaway E. G. Nucleotide and complete amino acid sequences of Kunjin virus: definitive gene order and characteristics of the virus-specified proteins. J Gen Virol. 1988 Jan;69(Pt 1):1–21. doi: 10.1099/0022-1317-69-1-1. [DOI] [PubMed] [Google Scholar]
  8. Dalgarno L., Trent D. W., Strauss J. H., Rice C. M. Partial nucleotide sequence of the Murray Valley encephalitis virus genome. Comparison of the encoded polypeptides with yellow fever virus structural and non-structural proteins. J Mol Biol. 1986 Feb 5;187(3):309–323. doi: 10.1016/0022-2836(86)90435-3. [DOI] [PubMed] [Google Scholar]
  9. Deubel V., Kinney R. M., Trent D. W. Nucleotide sequence and deduced amino acid sequence of the nonstructural proteins of dengue type 2 virus, Jamaica genotype: comparative analysis of the full-length genome. Virology. 1988 Jul;165(1):234–244. doi: 10.1016/0042-6822(88)90677-0. [DOI] [PubMed] [Google Scholar]
  10. Falgout B., Chanock R., Lai C. J. Proper processing of dengue virus nonstructural glycoprotein NS1 requires the N-terminal hydrophobic signal sequence and the downstream nonstructural protein NS2a. J Virol. 1989 May;63(5):1852–1860. doi: 10.1128/jvi.63.5.1852-1860.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hahn Y. S., Galler R., Hunkapiller T., Dalrymple J. M., Strauss J. H., Strauss E. G. Nucleotide sequence of dengue 2 RNA and comparison of the encoded proteins with those of other flaviviruses. Virology. 1988 Jan;162(1):167–180. doi: 10.1016/0042-6822(88)90406-0. [DOI] [PubMed] [Google Scholar]
  12. Kräusslich H. G., Wimmer E. Viral proteinases. Annu Rev Biochem. 1988;57:701–754. doi: 10.1146/annurev.bi.57.070188.003413. [DOI] [PubMed] [Google Scholar]
  13. Mackow E., Makino Y., Zhao B. T., Zhang Y. M., Markoff L., Buckler-White A., Guiler M., Chanock R., Lai C. J. The nucleotide sequence of dengue type 4 virus: analysis of genes coding for nonstructural proteins. Virology. 1987 Aug;159(2):217–228. doi: 10.1016/0042-6822(87)90458-2. [DOI] [PubMed] [Google Scholar]
  14. Mandl C. W., Heinz F. X., Stöckl E., Kunz C. Genome sequence of tick-borne encephalitis virus (Western subtype) and comparative analysis of nonstructural proteins with other flaviviruses. Virology. 1989 Nov;173(1):291–301. doi: 10.1016/0042-6822(89)90246-8. [DOI] [PubMed] [Google Scholar]
  15. Markoff L. In vitro processing of dengue virus structural proteins: cleavage of the pre-membrane protein. J Virol. 1989 Aug;63(8):3345–3352. doi: 10.1128/jvi.63.8.3345-3352.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mason P. W., McAda P. C., Mason T. L., Fournier M. J. Sequence of the dengue-1 virus genome in the region encoding the three structural proteins and the major nonstructural protein NS1. Virology. 1987 Nov;161(1):262–267. doi: 10.1016/0042-6822(87)90196-6. [DOI] [PubMed] [Google Scholar]
  17. Pletnev A. G., Yamshchikov V. F., Blinov V. M. Nucleotide sequence of the genome and complete amino acid sequence of the polyprotein of tick-borne encephalitis virus. Virology. 1990 Jan;174(1):250–263. doi: 10.1016/0042-6822(90)90073-z. [DOI] [PubMed] [Google Scholar]
  18. Rice C. M., Lenches E. M., Eddy S. R., Shin S. J., Sheets R. L., Strauss J. H. Nucleotide sequence of yellow fever virus: implications for flavivirus gene expression and evolution. Science. 1985 Aug 23;229(4715):726–733. doi: 10.1126/science.4023707. [DOI] [PubMed] [Google Scholar]
  19. Ruiz-Linares A., Cahour A., Després P., Girard M., Bouloy M. Processing of yellow fever virus polyprotein: role of cellular proteases in maturation of the structural proteins. J Virol. 1989 Oct;63(10):4199–4209. doi: 10.1128/jvi.63.10.4199-4209.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Speight G., Coia G., Parker M. D., Westaway E. G. Gene mapping and positive identification of the non-structural proteins NS2A, NS2B, NS3, NS4B and NS5 of the flavivirus Kunjin and their cleavage sites. J Gen Virol. 1988 Jan;69(Pt 1):23–34. doi: 10.1099/0022-1317-69-1-23. [DOI] [PubMed] [Google Scholar]
  21. Sumiyoshi H., Mori C., Fuke I., Morita K., Kuhara S., Kondou J., Kikuchi Y., Nagamatu H., Igarashi A. Complete nucleotide sequence of the Japanese encephalitis virus genome RNA. Virology. 1987 Dec;161(2):497–510. doi: 10.1016/0042-6822(87)90144-9. [DOI] [PubMed] [Google Scholar]
  22. Toyoda H., Nicklin M. J., Murray M. G., Anderson C. W., Dunn J. J., Studier F. W., Wimmer E. A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein. Cell. 1986 Jun 6;45(5):761–770. doi: 10.1016/0092-8674(86)90790-7. [DOI] [PubMed] [Google Scholar]
  23. Trent D. W., Kinney R. M., Johnson B. J., Vorndam A. V., Grant J. A., Deubel V., Rice C. M., Hahn C. Partial nucleotide sequence of St. Louis encephalitis virus RNA: structural proteins, NS1, ns2a, and ns2b. Virology. 1987 Feb;156(2):293–304. doi: 10.1016/0042-6822(87)90409-0. [DOI] [PubMed] [Google Scholar]
  24. Wengler G., Castle E., Leidner U., Nowak T., Wengler G. Sequence analysis of the membrane protein V3 of the flavivirus West Nile virus and of its gene. Virology. 1985 Dec;147(2):264–274. doi: 10.1016/0042-6822(85)90129-1. [DOI] [PubMed] [Google Scholar]
  25. Wright P. J., Cauchi M. R., Ng M. L. Definition of the carboxy termini of the three glycoproteins specified by dengue virus type 2. Virology. 1989 Jul;171(1):61–67. doi: 10.1016/0042-6822(89)90510-2. [DOI] [PubMed] [Google Scholar]
  26. Yaegashi T., Vakharia V. N., Page K., Sasaguri Y., Feighny R., Padmanabhan R. Partial sequence analysis of cloned dengue virus type 2 genome. Gene. 1986;46(2-3):257–267. doi: 10.1016/0378-1119(86)90410-5. [DOI] [PubMed] [Google Scholar]
  27. Zhao B. T., Prince G., Horswood R., Eckels K., Summers P., Chanock R., Lai C. J. Expression of dengue virus structural proteins and nonstructural protein NS1 by a recombinant vaccinia virus. J Virol. 1987 Dec;61(12):4019–4022. doi: 10.1128/jvi.61.12.4019-4022.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zhao B., Mackow E., Buckler-White A., Markoff L., Chanock R. M., Lai C. J., Makino Y. Cloning full-length dengue type 4 viral DNA sequences: analysis of genes coding for structural proteins. Virology. 1986 Nov;155(1):77–88. doi: 10.1016/0042-6822(86)90169-8. [DOI] [PubMed] [Google Scholar]
  29. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]

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