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
Article

Abstract

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

S944.41 

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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

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