Virologica Sinica

, Volume 24, Issue 1, pp 71–76 | Cite as

Identification and characterization of nuclear localization signals within the nucleocapsid protein VP15 of white spot syndrome virus

  • Li-juan Li
  • Hua-jun Zhang
  • Cong Zhang
  • Zheng-li Shi


The nucleocapsid protein VP15 of white spot syndrome virus (WSSV) is a basic DNA-binding protein. Three canonical bipartite nuclear localization signals (NLSs), called NLS1 (aa 11–27), NLS2 (aa 33–49) and NLS3 (44–60), have been detected in this protein, using the ScanProsite computer program. To determine the nuclear localization sequence of VP15, the full-length open reading frame, or the sequence of one of the three NLSs, was fused to the green fluorescent protein (GFP) gene, and transiently expressed in insect Sf9 cells. Transfection with full-length VP15 resulted in GFP fluorescence being distributed exclusively in the nucleus. NLS1 alone could also direct GFP to the nucleus, but less efficiently. Neither of the other two NLSs (NLS2 and 3) was functional when expressed alone, but exhibited similar activity to NLS1 when they were expressed as a fusion peptide. Furthermore, a mutated VP15, in which the two basic amino acids (11RR12) of NLS1 were changed to two alanines (11AA12), caused GFP to be localized only in the cytoplasm of Sf9 cells. These results demonstrated that VP15, as a nuclear localization protein, needs cooperation between its three NLSs, and that the two residues (11RR12) of NLS1 play a key role in transporting the protein to the nucleus.

Key words

White spot syndrome virus (WSSV) Nucleocapsid protein VP15 Nuclear localization signal (NLS) 

CLC number



Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Chen L L, Leu J H, Huang C J, et al. 2002. Identification of a nucleocapsid protein (VP35) gene of shrimp white spot syndrome virus and characterization of the motif important for targeting VP35 to the nuclei of transfected insect cells. Virology, 293(1): 44–53.PubMedCrossRefGoogle Scholar
  2. 2.
    Cokol M, Nair R, Rost B. 2000. Finding nuclear localization signals. EMBO Rep, 1(5): 411–415.PubMedCrossRefGoogle Scholar
  3. 3.
    Durand S, Lightner D V, Redman R M, et al. 1997. Ultrastructure and morphogenesis of white spot syndrome baculovirus (WSSV). Dis Aquat Organ, 29: 205–211.CrossRefGoogle Scholar
  4. 4.
    Gattiker A, Gasteiger E, Bairoch A. 2002. ScanProsite: a reference implementation of a PROSITE scanning tool. Appl Bioinformatics, 1(2): 107–108.PubMedGoogle Scholar
  5. 5.
    Hivin P, Frederic M, Arpin-Andre C, et al. 2005. Nuclear localization of HTLV-I bZIP factor (HBZ) is mediated by three distinct motifs. J Cell Sci, 118(Pt 7): 1355–1362.PubMedCrossRefGoogle Scholar
  6. 6.
    Hurt E C. 1993. The nuclear pore complex. FEBS Lett, 325(1–2): 76–80.PubMedCrossRefGoogle Scholar
  7. 7.
    Kalderon D, Roberts B L, Richardson W D, et al. 1984. A short amino acid sequence able to specify nuclear location. Cell, 39(3 Pt 2): 499–509.PubMedCrossRefGoogle Scholar
  8. 8.
    Kasamatsu H, Nakanishi A. 1998. How do animal DNA viruses get to the nucleus? Annu Rev Microbiol, 52: 627–686.PubMedCrossRefGoogle Scholar
  9. 9.
    Leu J H, Tsai J M, Wang H C, et al. 2005. The unique stacked rings in the nucleocapsid of the white spot syndrome virus virion are formed by the major structural protein VP664, the largest viral structural protein ever found. J Virol, 79(1): 140–149.PubMedCrossRefGoogle Scholar
  10. 10.
    Li Z, Lin Q, Chen J, et al. 2007. Shotgun identification of the structural proteome of shrimp white spot syndrome virus and iTRAQ differentiation of envelope and nucleocapsid subproteomes. Mol Cell Proteomics, 6(9): 1609–1620.PubMedCrossRefGoogle Scholar
  11. 11.
    Mayo M A. 2002. A summary of taxonomic changes recently approved by ICTV. Arch Virol, 147(8): 1655–1663.PubMedCrossRefGoogle Scholar
  12. 12.
    Robbins J, Dilworth S M, Laskey R A, et al. 1991. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell, 64(3): 615–623.PubMedCrossRefGoogle Scholar
  13. 13.
    Takahashi Y, Itami T, Kondo M, et al. 1994. Electron microscopy evidence of bacilliform virus infection in Kuruma shrimp (Penaeus japonicus). Fish Pathology, 29(2): 121–125.Google Scholar
  14. 14.
    Tsai J M, Wang H C, Leu J H, et al. 2004. Genomic and proteomic analysis of thirty-nine structural proteins of shrimp white spot syndrome virus. J Virol, 78(20): 11360–11370.PubMedCrossRefGoogle Scholar
  15. 15.
    Tsai J M, Wang H C, Leu J H, et al. 2006. Identification of the nucleocapsid, tegument, and envelope proteins of the shrimp white spot syndrome virus virion. J Virol, 80(6): 3021–3029.PubMedCrossRefGoogle Scholar
  16. 16.
    van Hulten M C, Reijns M, Vermeesch A M, et al. 2002. Identification of VP19 and VP15 of white spot syndrome virus (WSSV) and glycosylation status of the WSSV major structural proteins. J Gen Virol, 83(Pt 1):257–265.PubMedGoogle Scholar
  17. 17.
    van Hulten M C, Witteveldt J, Peters S, et al. 2001. The white spot syndrome virus DNA genome sequence. Virology, 286(1): 7–22.PubMedCrossRefGoogle Scholar
  18. 18.
    van Hulten M C W, Witteveldt J, Peters S, et al. 2001. The White Spot Syndrome Virus DNA Genome Sequence. Virology, 286(1): 7–22.PubMedCrossRefGoogle Scholar
  19. 19.
    Whittaker G R, Kann M, Helenius A. 2000. Viral entry into the nucleus. Annu Rev Cell Dev Biol, 16: 627–651.PubMedCrossRefGoogle Scholar
  20. 20.
    Witteveldt J, Vermeesch A M, Langenhof M, et al. 2005. Nucleocapsid protein VP15 is the basic DNA binding protein of white spot syndrome virus of shrimp. Arch Virol, 150(6): 1121–1333.PubMedCrossRefGoogle Scholar
  21. 21.
    Xie X, Xu L, Yang F. 2006. Proteomic analysis of the major envelope and nucleocapsid proteins of white spot syndrome virus. J Virol, 80(21): 10615–10623.PubMedCrossRefGoogle Scholar
  22. 22.
    Yang F, He J, Lin X, et al. 2001. Complete genome sequence of the shrimp white spot bacilliform virus. J Virol, 75(23): 11811–11820.PubMedCrossRefGoogle Scholar
  23. 23.
    Zhang X, Xu X, and Hew C L. 2001. The structure and function of a gene encoding a basic peptide from prawn white spot syndrome virus. Virus Res, 79(1–2): 137–144.PubMedCrossRefGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS and Springer-Verlag GmbH 2009

Authors and Affiliations

  • Li-juan Li
    • 1
    • 2
  • Hua-jun Zhang
    • 1
  • Cong Zhang
    • 1
  • Zheng-li Shi
    • 1
  1. 1.State Key Laboratory of VirologyWuhan Institute of Virology, Chinese Academy of SciencesWuhanChina
  2. 2.College of FisheryHuazhong Agricultural UniversityWuhanChina

Personalised recommendations