대한수의학회지 (Korean Journal of Veterinary Research)

  • 제45권2호
  • /
  • Pages.207-214
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  • 2005
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  • 2466-1384(pISSN)
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  • 2466-1392(eISSN)
대한수의학회 (The Korean Society of Veterinary Science)
권호 표지

조류 로타바이러스의 NSP4 유전자 염기서열분석 및 발현

Nucleotide sequence analysis and expression of NSP4 gene of avian rotavirus

  • 신인호 (충북대학교 수의과대학 동물의학연구소) ;
  • 이승철 (충북대학교 수의과대학 동물의학연구소) ;
  • 김원용 (중앙대학교 의과대학) ;
  • 강신영 (충북대학교 수의과대학 동물의학연구소)
  • Shin, In-ho (Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Lee, Seung-chul (Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University) ;
  • Kim, Won-yong (College of Medicine, Chungang University) ;
  • Kang, Shien-young (Research Institute of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University)
  • 심사 : 2005.05.27
  • 발행 : 2005.06.30
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초록

The nonstructural protein 4 (NSP4) of rotavirus encoded by gene 10, plays an important role in rotavirus pathogenicity. In this study, NSP4 gene of avian rotavirus (AvRV-1, AvRV-2) was analyzed and expressed using baculovirus expression system. The sequence data indicated that the NSP4 gene of AvRV-1 and AvRV-2 were 727 bases in length, encoded one open reading frame of 169 amino acids beginning at base 41 and terminating at base 550, and had two glycosylation sites. Nucleotide sequences of NSP4 gene of AvRV-1 and AvRV-2 exhibited a high degree of homology ($88.1{\pm}7.6%$) with avian rotaviruses, namely Ty1, Ty3 and PO-13. Phylogenetic analysis showed that AvRV-1 and AvRV-2 belonged to genotype NSP4[E], which is widely found in group A avian rotaviruses. The baculovirus-expressed NSP4 migrated at 20-28 kDa and reacted with NSP4-specific antiserum by FA and Western blot. Furthermore, it was found to be a glycoprotein by using tunicamycin, which is a specific inhibitor of N-linked glycosylation.

키워드

참고문헌

  1. 강신영, Nagaraja KV, Newman JA. Group A 조류 로타바이러스의 병원성에 관한 연구. 대한수의학회지 1993, 33, 217-225
  2. 백명순, 강신영, 김원용. 국내분리 소 로타바이러스 NSP4 유전자의 크로닝 및 염기 서열 분석. J Bacteriol Virol 2001, 31, 55-61
  3. Ball JM, Tian P, Zeng CQY, Morris AP, Estes MK. Age-dependent diarrhea induced by a rotaviral nonstructural glycoprotein. Science 1996, 272, 101-104 https://doi.org/10.1126/science.272.5258.101
  4. Bergeland ME, McAdaragh JP, Stotz I. Rotaviral enteritis in turkey poults. In: Proceedings of the 26th West Poultry Disease Conference. p.129-130, 1977
  5. Bridger JC. Novel rotaviruses in animals and man. Ciba Found Symp 1987, 128, 5-23
  6. Burke BM, McCrae A, Desselberger U. Sequence analysis of two porcine rotaviruses differing in growth in vitro and in pathogenicity: distinct VP4 sequences and conservation of NS53, VP6 and VP7 genes. J Gen Virol 1994, 75, 2205-2212 https://doi.org/10.1099/0022-1317-75-9-2205
  7. Chowdhury K. One step 'miniprep' method for the isolation of plasmid DNA. Nucleic Acids Res 1991, 19, 2792 https://doi.org/10.1093/nar/19.10.2792
  8. Ciarlet M, Liprandi F, Conner ME, Estes MK. Species specificity and interspecies relatedness of NSP4 genetic groups by comparative NSP4 sequence analyses of animal rotaviruses. Arch Virol 2000, 145, 371-383 https://doi.org/10.1007/s007050050029
  9. Davitt CM, Reynolds DL. Characterization of a group D rotavirus. Avian Dis 1993, 37, 749-755 https://doi.org/10.2307/1592024
  10. Dong YJ, Zeng CQY, Ball JM, Estes MK, Morris AP. The rotavirus enterotoxin NSP4 mobilizes intracellular calcium in human intestinal cells by stimulating phospholipase C mediated inositol 1,4,5-triphosphate production. Proc Natl Acad Sci USA 1997, 94, 3960-3965
  11. Estes MK, Cohen J. Rotavirus gene structure and function. Microbiol Rev 1989, 53, 410-449
  12. Estes MK, Palmer EL, Obijeski JF. Rotaviruses: a review. Curr Top Microbiol Immunol 1983, 105, 123-184
  13. Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 1985, 39, 783-791 https://doi.org/10.2307/2408678
  14. Felsenstein J. PHYLIP (Phylogeny Inference Package) version 3.5c. Department of Genetics, University of Washington, Seattle, 1993
  15. Ito H, Sugiyama M, Masubuchi K, Mori Y, Minamoto N. Complete nucleotide sequence of a group A avian rotavirus genome and a comparison with its counterparts of mammalian rotaviruses. Virus Res 2001, 75, 123-138 https://doi.org/10.1016/S0168-1702(01)00234-9
  16. Jukes TH, Cantor CR. Evolution of protein molecules. In: Munro HN (ed.). Mammalian protein metabolism, Vol. 3. pp. 21-132, Academic Press Inc, New York, 1969
  17. Kang SY, Saif LJ. Production and characterization of monoclonal antibodies against an avian group A rotavirus. Avian Dis 1991, 35, 563-571 https://doi.org/10.2307/1591221
  18. Lin SL, Tian P. Detailed computational analysis of a comprehensive set of group A rotavirus NSP4 proteins. Virus Genes 2003, 26, 271-282 https://doi.org/10.1023/A:1024451314534
  19. McNulty MS, Todd D, Allan GM, McFerran JB, Greene JA. Epidemiology of rotavirus infection in broiler chicken: recognition of four serogroups. Arch Virol 1984, 81, 113-121 https://doi.org/10.1007/BF01309301
  20. Mori Y, Borgan MA, Ito N, Sugiyama M, Minamoto N. Sequential analysis of nonstructural protein NSP4s derived from group A avian rotaviruses. Virus Res 2002, 89, 145-151 https://doi.org/10.1016/S0168-1702(02)00112-0
  21. Pedley S, Bridger JC, Chasey D, McCrae MA. Definition of two new groups of atypical rotaviruses. J Gen Virol 1986, 67, 131-137 https://doi.org/10.1099/0022-1317-67-1-131
  22. Petrie BI, Estes MK, Graham DY. Effects of tunicamycin on rotavirus morphogenesis and infectivity. J Virol 1983, 46, 270-274
  23. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987, 4, 406-425
  24. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X Windows Interface: flexible strategies for multiple sequence alignment aided by quality analysis tool. Nucleic Acid Res 1997, 24, 4876-4882
  25. Tian P, Estes MK, Hu Y, Ball JM, Zeng CQY, Schilling WP. The rotaviral nonstructural glycoprotein NSP4 mobilizes $Ca^{2+}$ from the endoplasmic reticulum. J Virol 1995, 69, 5763-5772
  26. Tian P, Hu Y, Schilling WP, Lindsay DA, Eiden J, Estes MK. The nonstructural glycoprotein of rotavirus affects intracellular calcium levels. J Virol 1994, 68, 251-257
  27. Zhang M, Zeng CQY, Dong Y, Ball JM, Saif LJ, Morris AP, Estes MK. Mutations in rotavirus nonstructural glycoprotein NSP4 are associated with altered virus virulence. J Virol 1998, 72, 3666-3672
  28. Zhang M, Zeng CQY, Morris AP, Estes MK. Functional NSP4 enterotoxin peptide secreted from rotavirus-infected cells. J Virol 2000, 74, 11663-11670 https://doi.org/10.1128/JVI.74.24.11663-11670.2000