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Functional expression of the recombinant ATPase of orf virus

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Abstract

Nucleotide sequence analysis has indicated that the A32L gene of orf virus can encode an ATPase (Chan et al. in Gene 432:44–53, 2009). In this work, we cloned the A32L gene into a prokaryotic expression vector, and the recombinant protein was expressed and purified. The antigenicity of recombinant ATPase was examined by immunoblotting, and its identity was confirmed by mass spectrometry. The ATP hydrolysis function of the purified recombinant protein was examined, and our results showed that it exhibited the ATPase activity. Similar to other viral ATPases, the ATPase of orf virus remained active in the presence of different divalent ions; nevertheless, unlike other viral ATPases, our recombinant ATPase exhibited similar enzymatic activity in reaction buffers of different pH.

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References

  1. Bayliss CD, Condit RC (1995) The vaccinia virus A18R gene product is a DNA-dependent ATPase. J Biol Chem 270:1550–1556

    Article  CAS  PubMed  Google Scholar 

  2. Cassetti MC, Merchlinsky M, Wolffe EJ, Weisberg AS, Moss B (1998) DNA packaging mutant: repression of the vaccinia virus A32 gene results in noninfectious, DNA-deficient, spherical, enveloped particles. J Virol 72:5769–5780

    CAS  PubMed  Google Scholar 

  3. Chan KW, Hsu WL, Wang CY, Yang CH, Lin FY, Chulakasian S, Wong ML (2009) Differential diagnosis of orf viruses by a single-step PCR. J Virol Methods 160:85–89

    Article  CAS  PubMed  Google Scholar 

  4. Chan KW, Yang CH, Lin JW, Wang HC, Lin FY, Kuo ST, Wong ML, Hsu WL (2009) Phylogenetic analysis of parapoxviruses and the C-terminal heterogeneity of viral ATPase proteins. Gene 432:44–53

    Article  CAS  PubMed  Google Scholar 

  5. Damon IK (2007) Poxviruses. In: Knipe DM, Griffin DE, Martin MA, Lamb RA, Roizman B, Straus SE (eds) Fields virology. Lippincott, Williams and Wilkins, Philadelphia, pp 2947–2975

    Google Scholar 

  6. Delhon G, Tulman ER, Afonso CL, Lu Z, de la Concha-Bermejillo A, Lehmkuhl HD, Piccone ME, Kutish GF, Rock DL (2004) Genomes of the parapoxviruses ORF virus and bovine papular stomatitis virus. J Virol 78:168–177

    Article  CAS  PubMed  Google Scholar 

  7. Deng L, Shuman S (1998) Vaccinia NPH-I, a DExH-box ATPase, is the energy coupling factor for mRNA transcription termination. Genes Dev 12:538–546

    Article  CAS  PubMed  Google Scholar 

  8. Gumbrell RC, McGregor DA (1997) Outbreak of severe fatal orf in lambs. Vet Rec 141:150–151

    CAS  PubMed  Google Scholar 

  9. Haig DM, Mercer AA (1998) Ovine diseases. Orf Vet Res 29:311–326

    CAS  Google Scholar 

  10. Iyer LM, Aravind L, Koonin EV (2001) Common origin of four diverse families of large eukaryotic DNA viruses. J Virol 75:11720–11734

    Article  CAS  PubMed  Google Scholar 

  11. Koonin EV, Senkevich TG, Chernos VI (1993) Gene A32 product of vaccinia virus may be an ATPase involved in viral DNA packaging as indicated by sequence comparisons with other putative viral ATPases. Virus Genes 7:89–94

    Article  CAS  PubMed  Google Scholar 

  12. Kuo MD, Chin C, Hsu SL, Shiao JY, Wang TM, Lin JH (1996) Characterization of the NTPase activity of Japanese encephalitis virus NS3 protein. J Gen Virol 77(Pt 9):2077–2084

    Article  CAS  PubMed  Google Scholar 

  13. Mazur C, Machado RD (1989) Detection of contagious pustular dermatitis virus of goats in a severe outbreak. Vet Rec 125:419–420

    CAS  PubMed  Google Scholar 

  14. Mercer AA, Ueda N, Friederichs SM, Hofmann K, Fraser KM, Bateman T, Fleming SB (2006) Comparative analysis of genome sequences of three isolates of Orf virus reveals unexpected sequence variation. Virus Res 116:146–158

    CAS  PubMed  Google Scholar 

  15. Mondal B, Bera AK, Hosamani M, Tembhurne PA, Bandyopadhyay SK (2006) Detection of Orf virus from an outbreak in goats and its genetic relation with other parapoxviruses. Vet Res Commun 30:531–539

    Article  CAS  PubMed  Google Scholar 

  16. Moss B (2007) Poxviridae: the viruses and their replication. In: Knipe DM, Griffin DE, Martin MA, Lamb RA, Roizman B, Straus SE (eds) Fields virology. Lippincott, Williams and Wilkins, Philadelphia, pp 2905–2945

    Google Scholar 

  17. Ramadevi N, Roy P (1998) Bluetongue virus core protein VP4 has nucleoside triphosphate phosphohydrolase activity. J Gen Virol 79(Pt 10):2475–2480

    CAS  PubMed  Google Scholar 

  18. Reid HW (1991) Disease of sheep, 2nd edn. Oxford, Blackwell scientific publication, London

    Google Scholar 

  19. Simpson DA, Condit RC (1995) Vaccinia virus gene A18R encodes an essential DNA helicase. J Virol 69:6131–6139

    CAS  PubMed  Google Scholar 

  20. Su YP, Shien JH, Liu HJ, Yin HS, Lee LH (2007) Avian reovirus core protein muA expressed in Escherichia coli possesses both NTPase and RTPase activities. J Gen Virol 88:1797–1805

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan

    Fong-Yuan Lin, Hsien-Chi Wang & Min-Liang Wong

  2. Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan

    Kun-Wei Chan

  3. Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan

    Wei-Li Hsu

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  1. Fong-Yuan Lin
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  2. Kun-Wei Chan
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  3. Hsien-Chi Wang
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  4. Wei-Li Hsu
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  5. Min-Liang Wong
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Correspondence to Wei-Li Hsu or Min-Liang Wong.

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Lin, FY., Chan, KW., Wang, HC. et al. Functional expression of the recombinant ATPase of orf virus. Arch Virol 155, 1701–1705 (2010). https://doi.org/10.1007/s00705-010-0754-5

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