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

Virologica Sinica volume 24, pages 71–76 (2009)Cite this 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.

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Authors and Affiliations

  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China

    Li-juan Li, Hua-jun Zhang, Cong Zhang & Zheng-li Shi

  2. College of Fishery, Huazhong Agricultural University, Wuhan, 430070, China

    Li-juan Li

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  1. Li-juan Li
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  2. Hua-jun Zhang
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  3. Cong Zhang
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  4. Zheng-li Shi
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Correspondence to Zheng-li Shi.

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Foundation item: State Key Program for Basic Research Grants (2006CB101801); the Chinese Academy of Sciences (KSCX2-SW-302).

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Li, Lj., Zhang, Hj., Zhang, C. et al. Identification and characterization of nuclear localization signals within the nucleocapsid protein VP15 of white spot syndrome virus. Virol. Sin. 24, 71–76 (2009). https://doi.org/10.1007/s12250-009-3013-8

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  • DOI: https://doi.org/10.1007/s12250-009-3013-8

Key words

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

CLC number

  • S944.41