Skip to main content
SpringerLink
Log in

Identification of a novel bipartite nuclear localization signal in the DNA polymerase of the betabaculovirus Pieris rapae granulovirus

  • Brief Report
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

DNA polymerase (DNApol) is highly conserved in all baculoviruses and plays an essential role in viral DNA replication. Previous results showed that the DNApol of the betabaculovirus Pieris rapae granulovirus (PiraGV) can localize in the nucleus. However, it is not clear how the DNApol is transported into the nucleus. Bioinformatic and GFP localization analysis showed that PiraGV DNApol contains a nuclear localization signal (NLS) at aa 4-25 (LFKRKLDEPPTDHTLVKAIKLS) of the N-terminus that does not match either the classical monopartite or the bipartite NLS consensus sequence. Multiple-point-substitution analysis confirmed that the NLS is required for transport of PiraGV DNApol into the nucleus. We also substituted the NLS of the PiraGV DNApol for that of the alphabaculovirus Spodoptera litura nuclear polyhedrosis virus (SpltNPV) DNApol. A viral growth curve and quantitative real-time PCR revealed that the substitution impaired viral DNA replication and resulted in a reduction in virus production. Together, our results show that PiraGV contains a novel NLS and that the NLS cannot efficiently replace that of SpltNPV DNApol for viral DNA synthesis and virus production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

References

  1. Au V, Yu M, Carstens EB (2009) Characterization of a baculovirus nuclear localization signal domain in the late expression factor 3 protein. Virology 385:209–217

    Article  CAS  PubMed  Google Scholar 

  2. Basil A, Shannon E, Lillian P (2011) Biology and genomics of viruses within the genus Gammabaculovirus. Viruses 3:2214

    Article  CAS  Google Scholar 

  3. Braithwaite DK, Ito J (1993) Compilation, alignment, and phylogenetic relationships of DNA polymerases. Nucleic Acids Res 21:787–802

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Chen GQ, Fang Y, Hu ZY, Krell PJ, Feng GZ (2016) Rescue of dnapol-null Autographa californica multiple nucleopolyhedrovirus with DNA polymerase (DNApol) of Spodoptera litura nucleopolyhedrovirus (SpltNPV) and identification of a nuclear localization signal in SpltNPV DNApol. J Gen Virol 97:1968–1980

    Article  CAS  PubMed  Google Scholar 

  5. Chen GQ, Fang Y, Wu LJ, Yan Q, Krell PJ, Feng GZ (2017) A betabaculovirus DNA polymerase cannot substitute for the DNA polymerase of the alphabaculovirus Autographa californica nucleopolyhedrovirus. Arch Virol 162:3487–3492

    Article  CAS  PubMed  Google Scholar 

  6. Chen ZL, Carstens EB (2005) Identification of domains in Autographa californica multiple nucleopolyhedrovirus late expression factor 3 required for nuclear transport of P143. J Virol 79:10915–10922

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Clem RJ, Passarelli AL (2013) Baculoviruses: sophisticated pathogens of insects. PLoS Pathog 9:e1003729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Feng GZ, Krell PJ (2014) Autographa californica multiple nucleopolyhedrovirus DNA polymerase C terminus is required for nuclear localization and viral DNA replication. J Virol 88:10918–10933

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Friedrich B, Quensel C, Sommer T, Hartmann E, Kohler M (2006) Nuclear localization signal and protein context both mediate importin at specificity of nuclear import substrates. Mol Cell Biol 26:8697–8709

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Gorlich D, Kutay U (1999) Transport between the cell nucleus and the cytoplasm. Annu Rev Cell Dev Biol 15:607–660

    Article  CAS  PubMed  Google Scholar 

  11. Guo ZJ, Wang DX, Yao Q, Chen KP, Zhang CX (2010) Identification of a novel functional nuclear localization signal in the protein encoded by open reading frame 47 of Bombyx mori nucleopolyhedrovirus. Arch Virol 155:1943–1950

    Article  CAS  PubMed  Google Scholar 

  12. Hang X, Guarino LA (1999) Purification of Autographa californica nucleopolyhedrovirus DNA polymerase from infected insect cells. J Gen Virol 80:2519–2526

    Article  CAS  PubMed  Google Scholar 

  13. Holt JE, Ly-Huynh JD, Efthymiadis A, Hime GR, Loveland KL, Jans DA (2007) Regulation of nuclear import during differentiation; The IMP alpha gene family and spermatogenesis. Curr Genom 8:323–334

    Article  CAS  Google Scholar 

  14. Huang J, Levin DB (2001) Identification, transcription and sequence analysis of the Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) DNA polymerase gene. Arch Virol 146:303–326

    Article  CAS  PubMed  Google Scholar 

  15. Hwang YT, Zuccola HJ, Lu Q, Hwang CB (2004) A point mutation within conserved region VI of herpes simplex virus type 1 DNA polymerase confers altered drug sensitivity and enhances replication fidelity. J Virol 78:650

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Jarvis DL, Bohlmeyer DA, Garcia A Jr (1991) Requirements for nuclear localization and supramolecular assembly of a baculovirus polyhedrin protein. Virology 185:795–810

    Article  CAS  PubMed  Google Scholar 

  17. Jehle JA, Blissard GW, Bonning BC, Cory JS, Herniou EA, Rohrmann GF, Theilmann DA, Thiem SM, Vlak JM (2006) On the classification and nomenclature of baculoviruses: a proposal for revision. Arch Virol 151:1257–1266

    Article  CAS  PubMed  Google Scholar 

  18. Jehle JA, Lange M, Wang H, Hu Z, Wang Y, Hauschild R (2006) Molecular identification and phylogenetic analysis of baculoviruses from Lepidoptera. Virology 346:180–193

    Article  CAS  PubMed  Google Scholar 

  19. Knopf CW (1998) Evolution of viral DNA-dependent DNA polymerases. Virus Genes 16:47–58

    Article  CAS  PubMed  Google Scholar 

  20. Kool M, Ahrens CH, Goldbach RW, Rohrmann GF, Vlak JM (1994) Identification of genes involved in DNA replication of the Autographa californica baculovirus. Proc Natl Acad Sci USA 91:11212–11216

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Kosugi S, Hasebe M, Matsumura N, Takashima H, Miyamoto-Sato E, Tomita M, Yanagawa H (2009) Six classes of nuclear localization signals specific to different binding grooves of importin alpha. J Biol Chem 284:478–485

    Article  CAS  PubMed  Google Scholar 

  22. Lange A, Mills RE, Lange CJ, Stewart M, Devine SE, Corbett AH (2007) Classical nuclear localization signals: definition, function, and interaction with importin alpha. J Biol Chem 282:5101–5105

    Article  CAS  PubMed  Google Scholar 

  23. Liu H, Naismith JH, Hay RT (2000) Identification of conserved residues contributing to the activities of adenovirus DNA polymerase. J Virol 74:11681–11689

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Lu A, Miller LK (1995) The roles of eighteen baculovirus late expression factor genes in transcription and DNA replication. J Virol 69:975–982

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Macara IG (2001) Transport into and out of the nucleus. Microbiol Mol Biol Rev 65:570–594

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. McDougal VV, Guarino LA (1999) Autographa californica nuclear polyhedrosis virus DNA polymerase: measurements of processivity and strand displacement. J Virol 73:4908–4918

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Olson VA, Wetter JA, Friesen PD (2002) Baculovirus transregulator IE1 requires a dimeric nuclear localization element for nuclear import and promoter activation. J Virol 76:9505–9515

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Tomalski MD, Wu JG, Miller LK (1988) The location, sequence, transcription, and regulation of a baculovirus DNA polymerase gene. Virology 167:591–600

    Article  CAS  PubMed  Google Scholar 

  29. Wang Y, Wang Q, Liang CY, Song JH, Li N, Shi H, Chen XW (2008) Autographa californica multiple nucleopolyhedrovirus nucleocapsid protein BV/ODV-C42 mediates the nuclear entry of P78/83. J Virol 82:4554–4561

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Weis K (2003) Regulating access to the genome: nucleocytoplasmic transport throughout the cell cycle. Cell 112:441–451

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This research was supported by the National Natural Science Foundation of the China (Grant nos. 31572006 and 31701847), innovation project of Chinese Academy of Agricultural Sciences, and the Central Level Public Interest Research Institute for Basic R & D Special Fund Business (Grant no. 2017RG002-7).

Author information

Authors and Affiliations

  1. College of Agriculture, Yangtze University, Jingzhou, Hubei, China

    Pei Li

  2. State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou, 311400, China

    Pei Li, Lijuan Wu & Guoqing Chen

Authors
  1. Pei Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lijuan Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guoqing Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guoqing Chen.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Handling Editor: T. K. Frey.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 443 kb)

Supplementary material 2 (DOCX 19 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, P., Wu, L. & Chen, G. Identification of a novel bipartite nuclear localization signal in the DNA polymerase of the betabaculovirus Pieris rapae granulovirus. Arch Virol 164, 839–845 (2019). https://doi.org/10.1007/s00705-018-4115-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-018-4115-0

Search

Navigation