Characterization of the v-cath Gene of Bombyx mori Nuclear Polyhedrosis Virus K1

  • Lee, Kwang Sik (College of Natural Resources and Life Science, Dong-A University) ;
  • Li, Jianhong (College of Natural Resources and Life Science, Dong-A University) ;
  • Je, Yeon Ho (School of Agricultural Biotechnology, Seoul National University) ;
  • Woo, Soo Dong (Department of Plant Medicine, Chungbuk National University) ;
  • Sohn, Hung Dae (College of Natural Resources and Life Science, Dong-A University) ;
  • Jin, Byung Rae (College of Natural Resources and Life Science, Dong-A University)
  • Published : 2004.12.01

Abstract

A cathepsin L-like cysteine protease, v-cath, encoded by the baculovirus has been shown to playa role in host liquefaction. We have identified a v-cath gene in the silkworm virus, Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 969 bp v-cath has an open reading frame of 323 amino acids. A putative cleavage site and catalytic sites were conserved in BmNPV-K1 v-cath. The predicted three-dimensional structure of BmNPV-K1 v-cath revealed that the overall fold of BmNPV-K1 v-cath is similar to that of other proteases of the papain family. The deduced amino acid sequence of BmNPV-K1 v-cath showed 98% and 97% protein sequence identity to BmNPV T3 strain and to Autographa californica nuclear polyhedrosis virus, respectively. The BmNPV-K1 v-cath differed at 4 amino acid positions from BmNPV T3. The v-cath gene in BmNPV-K1 genome is located on the EcoRV 6 kb and XhoI 9 kb fragments. Northern hybridization analysis of BmNPV K1 v-cath gene revealed that it is expressed late in infection.

Keywords

References

  1. Ahrens, C. H., R. L. Russell, C. J. Funk, J. T. Evans, S. H. Harwood and G. F. Rohrmann (1997) The sequence of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. Virology 229, 381-399 https://doi.org/10.1006/viro.1997.8448
  2. Ayres, M. D., S. C. Howard, J. Kuzio, M. Lopez-Ferber and R. D. Possee (1994) The complete DNA sequence of the Autographa californica nuclear polyhedrosis virus. Virology 202, 586-605 https://doi.org/10.1006/viro.1994.1380
  3. Gal, S. and M. M. Gottesman (1988) Isolation and sequence of a cDNA for human pro-(cathepsin L). Biochem. J. 253, 303-306 https://doi.org/10.1042/bj2530303
  4. Gomi, S., K. Majima and S. Maeda (1999) Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J. Gen. Virol. 80, 1323-1337 https://doi.org/10.1099/0022-1317-80-5-1323
  5. Grace, T. C. D. (1962) Establishment of four strains of cells from insect tissue grown in vitro. Nature 195, 788-789 https://doi.org/10.1038/195788a0
  6. Hawtin, R. E., T. Zarkowska, K. Arnold, C. J. Thomas, G. W. Gooday, L. A. King, J. A. Kuzio and R. D. Possee (1997) Liquefaction of Autographa californica nucleopolyhedrovirus-infected insect is dependent on the integrity of virusencoded chitinase and cathepsin genes. Virology 238, 243-253 https://doi.org/10.1006/viro.1997.8816
  7. Hill, J. E., J. Kuzio and P. Faulkner (1995) Identification and characterization of the v-cath gene of the baculovirus, CfM-NPV. Biochim. Biophys. Acta 1264, 275-278 https://doi.org/10.1016/0167-4781(95)00195-6
  8. Hom, L. G., T. Ohkawa, D. Trudeau and L. E. Volkman (2002) Autographa californica M nucleopolyhedrovirus ProV-CATH is activated during infected cell death. Virology 296, 212-218 https://doi.org/10.1006/viro.2002.1378
  9. Hom, L. G. and L. E. Volkman (2000) Autographa californica M nucleopolyhedrovirus chiA is required for processing of V-CATH. Virology 277, 178-183 https://doi.org/10.1006/viro.2000.0586
  10. Kang, S. W., B. R. Jin, E. Y. Yun, S. H. Kim, K. Y. Kim and S. K. Kang (1997) Construction of the novel baculovirus transfer vector using the p10 gene of BmNPV. Korean J. Seric. Sci. 39, 180-185
  11. King, L. A. and R. D. Possee (1992) The baculovirus expression system. A laboratory giude. Chapman & Hall. London
  12. Lanier, L. M., J. M. Slack and L. E. Volkman (1996) Actin binding and proteolysis by the baculovirus AcMNPV: The role of virion-associated V-CATH. Virology 216, 380-388 https://doi.org/10.1006/viro.1996.0073
  13. Lee, K. S., H. J. Park, Y. H. Je, H. D. Sohn and B. R. Jin (2001) Molecular cloning of the antiapoptotic gene, p35, from Bombyx mori nuclear polyhedrosis virus K1. Int. J. Indust. Entomol. 3, 25-29
  14. Maeda, S. (1984) A plaque assay and cloning of Bombyx mori nuclear polyhedrosis virus. J. Serie. Sci. Jpn 53, 547-548
  15. Maeda, S., T. Kawai, M. Obinata, H. Fujiwara, T. Horiuchi, Y. Saeki, Y. Sato and M. Furusawa (1985) Production of human alpha-interferon in silkworm using a baculovirus vector. Nature 315, 592-594 https://doi.org/10.1038/315592a0
  16. McMaster, G. K. and G. G. Carmichael (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc. Natl. Acad. Sci. USA 74, 4835-4838 https://doi.org/10.1073/pnas.74.11.4835
  17. Ohkawa, T., K. Majima and S. Maeda (1994) A cysteine proteinase encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus. J. Virol. 68, 6619-6625
  18. O'Reilly, D. R., L. K. Miller and V. A. Luckow (1992) Baculovirus expression vectors: a laboratory manual. W. H. Freeman & Co., New York
  19. Park, H. J., K. S. Lee, E. S. Cho, E. Y. Yun, S. W. Kang, K. Y. Kim, H. D. Sohn and B. R. Jin (2000) Molecular cloning and characterization of very late expression factor I gene, vlf-1 from Bombyx mori nuclear polyhedrosis virus K1. Int. J. Indust. Entomol. 1, 29-33
  20. Park, H. J., K. S. Lee, Y. H. Je, H. D. Sohn and B. R. Jin (2001a) Molecular cloning and sequence analysis of the immediate early I gene, lE1, from Bombyx mori nuclear polyhedrosis virus K1. Int. J. Indust. Entomol. 3, 43-49
  21. Park, H. J., E. H. Chung, K. S. Lee, J. H. Han, S. J. Lee, H. D. Sohn and B. R. Jin (2001b) Molecular cloning and sequencing of the ecdysteroid UDP-glucosyltransferase, EGT, from Bombyx mori nuclear polyhedrosis virus K1. Int. J. Indust. Entomol. 3, 37-41
  22. Rawlings, N. D., L. H. Pearl and D. J. Buttle (1992) The baculovirus Autographa californica nuclear polyhedrosis virus genome includes a papain-like sequence. Biol. Chem. Hoppe-Seyler 373, 1211-1215 https://doi.org/10.1515/bchm3.1992.373.2.1211
  23. Schwede, T., J. Kopp, N. Guex and M. C. Peitsch (2003) SWISS-MODEL: an automated protein homology-modeling server. Nucl. Acids Res. 31, 3381-3385 https://doi.org/10.1093/nar/gkg520
  24. Sivaraman, J., D. K. Nagler, R. Zhang, R. Menard and M. Cygler (2000) Crystal structure of human procathepsin X: A cysteine protease with the proregion covalently linked to the active site cysteine. J. Mol. Biol. 295, 939-951 https://doi.org/10.1006/jmbi.1999.3410
  25. Slack, J. M., J. Kuzio and P. Faulkner (1995) Characterization of v-cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. J. Gen. Virol. 76, 1091-1098 https://doi.org/10.1099/0022-1317-76-5-1091
  26. Vernett, T., D. C. Tessier, C. Richardson, F. Laliberte, H. E. Khouri, A. W. Bell, A. C. Storer and D. Y. Thomas (1990) Secretion of functional papain precursor from insect cells. Requirement for N-glycosylation of the pro-region. J. Biol. Chem. 265, 16661-16670
  27. Woo, S. D., W. J. Kim, B. R. Jin and S. K. Kang (1995) Construction of new transfer vector of nuclear polyhedrosis virus of the silkworm, Bombyx mori. Korean J. Seric. Sci. 37, 46-51