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Volume 25 Issue 2
April 2010
    Citation: Ling SHAO, Chao FAN, Ewa Maj, Qin FANG. Molecular Characterization of Nonstructural Protein NS38 of Grass Carp Reovirus* .VIROLOGICA SINICA, 2010, 25(2) : 123-129.  http://dx.doi.org/10.1007/s12250-010-3115-3
    shu

    Molecular Characterization of Nonstructural Protein NS38 of Grass Carp Reovirus*

    cstr: 32224.14.s12250-010-3115-3
    • 1.

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

    • 2.

      Graduate School of the Chinese Academy of Sciences, Beijing 100039, China

    • Corresponding author: Qin FANG, qfang@wh.iov.cn
    • † Current address: Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, ul. Kładki 24,80-822, Gdańsk, Poland
    • Received Date: 14 December 2009
      Accepted Date: 22 January 2010
      Available online: 01 April 2010

      Fund Project: National Basic Research Program (973) of China 2009CB118701National Natural Scientific Foundation of China 30671615National Natural Scientific Foundation of China 30871940

    • Viral nonstructural proteins in both enveloped and non-enveloped viruses play important roles in viral replication. Protein NS38 of Grass carp reovirus (GCRV), has been deduced to be a non-structural protein, and, consistent with other reoviruses, is considered to cooperate with the NS80 protein in viral particle assembly. To investigate the molecular basis of the role of NS38, a complete protein was expressed in E.coli for the first time. It was found that there is a better expression of NS38 induced with IPTG at 28 ℃ rather than 37 ℃. In addition, the antiserum of NS38 prepared with purified fusion protein and injected into rabbit could be used for detecting NS38 protein expression in GCRV infected cell lysate, while there is not any reaction crossed with purified virus particle, confirming NS38 is not a component of the viral structural protein. The result reported in this study will provide evidence for further viral protein-protein and protein-RNA interaction in dsRNA viruses replication.
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      1. Arnold M M, Murray K E, Nibert M L. 2008. Formation of the factory matrix is an important, though not a sufficient function of nonstructural protein μNS during reovirus infection. Virology, 375(2): 412-423.
          doi: 10.1016/j.virol.2008.02.024

      2. Attoui H, Fang Q, Jaafar F M, et al. 2002. Common evolutionary origin of aquareoviruses and orthoreoviruses revealed by genome characterization of Golden shiner reovirus, Grass carp reovirus, Striped bass reovirus and golden ide reovirus (genus Aquareovirus, family Reoviridae). J Gen Virol, 83: 1941-1951.
          doi: 10.1099/0022-1317-83-8-1941

      3. Becker M M, Goral M I, Hazelton P R, et al. 2001. Reovirus σNS protein is required for nucleation of viral assembly complexes and formation of viral inclusions. J Virol, 75(3): 1459-1475.
          doi: 10.1128/JVI.75.3.1459-1475.2001

      4. Becker M M, Peters T R, Dermody T S. 2003. Reovirus σNS and μNS proteins form cytoplasmic inclusion structures in the absence of viral infection. J Virol, 77(10): 5948-5963.
          doi: 10.1128/JVI.77.10.5948-5963.2003

      5. Broering T J, Kim J, Miller C L, et al. 2004. Reovirus nonstructural protein μNS recruits viral core surface proteins and entering core particles to factory-like inclusions. J Virol, 78(4): 1882-1892.
          doi: 10.1128/JVI.78.4.1882-1892.2004

      6. Cheng L P, Fang Q, Shah S, et al. 2008. Subnanometer-resolution structures of the grass carp reovirus core and virion. J Mol Biol, 382(1): 213-222.
          doi: 10.1016/j.jmb.2008.06.075

      7. Coffey C M, Sheh A, Kim I S, et al. 2006. Reovirus outer capsid protein μ1 induces apoptosis and associates with lipid droplets, endoplasmic reticulum, and mitochondria. J Virol, 80(17): 8422-8438.
          doi: 10.1128/JVI.02601-05

      8. Fan C, Fang Q. 2009. Functional Analyses of Mammalian Reovirus Nonstructural Protein μNS. Virologica Sinica, 24(1): 1-8.
          doi: 10.1007/s12250-009-3016-5

      9. Fan C, Zhang L L, Fang Q, et al. 2009. Expression and identification of inclusion forming-related domain of NS80 nonstructural protein of grass carp reovirus. Virologica Sinica, 24(3): 194-201.
          doi: 10.1007/s12250-009-3028-1

      10. Fang Q, Ke L H, Cai Y Q. 1989. Growth characterization and high titre culture of GCHV. Virologica Sinica,4(3): 314-319.

      11. Fang Q, Sanket S, Liang Y Y, et al. 2005. 3D reconstruction and capsid protein characterization of grass carp reovirus. Science in China Series C-Life Sciences, 48(6): 593-600.
          doi: 10.1360/062004-105

      12. Fang Q, Seng E K, Ding Q Q, et al. 2008. Characterization of infectious particles of grass carp reovirus by treatment with proteases. Arch Virol, 153(4): 675-682.
          doi: 10.1007/s00705-008-0048-3

      13. Gillian A L, Schmechel S C, Livny J, et al. 2000. Reovirus protein σNS binds in multiple copies to single-stranded RNA and shares properties with single-stranded DNA binding proteins. J Virol, 74(13): 5939-5948.
          doi: 10.1128/JVI.74.13.5939-5948.2000

      14. Gomatos P J, Prakash O, Stamatos N M. 1981. Small reovirus particle composed solely of σNS with specificity for binding different nucleic acids. J Virol, 39(1): 115-124.

      15. Jaafar F M, Goodwin A E, Belhouchet M, et al. 2008. Complete characterisation of the American grass carp reovirus genome (genus Aquareovirus: family Reoviridae) reveals an evolutionary link between aquareoviruses and coltiviruses. Virology, 373(2): 310-321.
          doi: 10.1016/j.virol.2007.12.006

      16. Joseph S, Russell D W. 2001. Molecular Cloning-A laboratory manual.Third edition. New York: Cold Spring Harbor Laboratory Press.

      17. Liemann S, Chandran K, Baker T S, et al. 2002. Structure of the reovirus membrane-penetration protein, μ1, in a complex with its protector protein, σ3. Cell, 108(2): 283-295.
          doi: 10.1016/S0092-8674(02)00612-8

      18. Odegard A L, Chandran K, Zhang X, et al. 2004. Putative autocleavage of outer capsid protein μ1, allowing release of myristoylated peptide μ1N during particle uncoating, is critical for cell entry by reovirus. J Virol, 78(16): 8732-8745.
          doi: 10.1128/JVI.78.16.8732-8745.2004

      19. Rangel A A, Rockemann D D, Hetrick F M, et al. 1999. Identification of grass carp haemorrhage virus as a new genogroup of aquareovirus. J Gen Virol, 80 (Pt 9): 2399-2402.

      20. Shaw A L, Samal S K, Subramanian K, et al. 1996. The structure of aquareovirus shows how the different geometries of the two layers of the capsid are reconciled to provide symmetrical interactions and stabilization. Structure, 4(8): 957-967.
          doi: 10.1016/S0969-2126(96)00102-5

      21. Touris-Otero F, Martinez-Costas J, Vakharia V N, et al. 2005. Characterization of the nucleic acid-binding activity of the avian reovirus non-structural protein σNS. J Gen Virol, 86: 1159-1169.
          doi: 10.1099/vir.0.80491-0

      22. Van Regenmortel M H V, Fauquet C M, Bishop D H L, et al. 2000.Virus Taxonomy-Seventh Report of the International Committee on Taxonomy of Viruses. SanDiego:Academic Press, California, USA.

      23. Zhang L L, Shen J Y, Fang Q, et al. 2008. High level expression of grass carp reovirus VP7 protein in prokaryotic cells. Virologica Sinica, 23(1): 57-62.
          doi: 10.1007/s12250-008-2923-1

      24. Zou G P, Fang Q. 2000. Study on replication and morphogenesis of the Grass Carp Reovirus (GCRV) in CIK cells. Virologica Sinica, 15(2): 188-192.

      25. Zuo W G, Qian H X, Xue Y F, et al. 1984. Establishment of grass carp kidney cell line (CIK). Fresh-water Fisheries(2): 38-39.

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      Molecular Characterization of Nonstructural Protein NS38 of Grass Carp Reovirus*

        Corresponding author: Qin FANG, qfang@wh.iov.cn
      • 1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
      • 2. Graduate School of the Chinese Academy of Sciences, Beijing 100039, China
      • Received Date: 14 December 2009
      • Accepted Date: 22 January 2010
      Fund Project:  National Basic Research Program (973) of China 2009CB118701National Natural Scientific Foundation of China 30671615National Natural Scientific Foundation of China 30871940

      Abstract: Viral nonstructural proteins in both enveloped and non-enveloped viruses play important roles in viral replication. Protein NS38 of Grass carp reovirus (GCRV), has been deduced to be a non-structural protein, and, consistent with other reoviruses, is considered to cooperate with the NS80 protein in viral particle assembly. To investigate the molecular basis of the role of NS38, a complete protein was expressed in E.coli for the first time. It was found that there is a better expression of NS38 induced with IPTG at 28 ℃ rather than 37 ℃. In addition, the antiserum of NS38 prepared with purified fusion protein and injected into rabbit could be used for detecting NS38 protein expression in GCRV infected cell lysate, while there is not any reaction crossed with purified virus particle, confirming NS38 is not a component of the viral structural protein. The result reported in this study will provide evidence for further viral protein-protein and protein-RNA interaction in dsRNA viruses replication.

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