Journal of Microbiology

  • 제44권5호
  • /
  • Pages.556-561
  • /
  • 2006
  • /
  • 1225-8873(pISSN)
  • /
  • 1976-3794(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Cloning and Characterization of a Thioredoxin Gene, CpTrx1, from the Chestnut Blight Fungus Cryphonectria parasitica

  • Kim, Ji-Hye (Basic Science Research Institute, Institute for Molecular Biology and Genetics, Chonbuk National University) ;
  • Kim, Dae-Hyuk (Basic Science Research Institute, Institute for Molecular Biology and Genetics, Chonbuk National University)
  • 발행 : 2006.10.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

A differential display for the expression profiles of wild-type Cryphonectria parasitica and its virally-infected isogenic hypovirulent strain revealed several transcripts of interest, which evidenced significant matches with fungal genes of known function. Among which, we have further analyzed an amplified PCR product with significant sequence similarity to the known fungal stress-responsive thioredoxin gene from Neurospora crassa. The product of the cloned thioredoxin gene, CpTrx1, consists of 117 amino acids, with a predicted molecular mass of 13.0 kDa and a pI of 5.4. Sequence comparisons demonstrated that the deduced protein sequence of the CpTrx1 gene evidenced a high degree of homology to all known thioredoxins, with the highest degree of homology with trx1, a thioredoxin gene from Saccharomyces cerevisiae, and evidenced a preservation of the conserved hall markresidues (Trp-Cys-Gly-Pro-Cys) at the active site of thioredoxin. The E. coli-generated CpTRX1 manifested thioredoxin activity, according to the insulin reduction assay, which indicates that the cloned gene does indeed encode for the C. parasitica thioredoxin.

키워드

참고문헌

  1. Allen, T.D., A.L. Dawe, and D.L. Nuss. 2003. Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonectria parasitica to infection by virulence-attenuating hypoviruses. Eukaryot. Cell 2, 1253-1265 https://doi.org/10.1128/EC.2.6.1253-1265.2003
  2. Allen, T.D. and D.L. Nuss. 2004. Specific and common alterations in host gene transcript accumulation following infection of the chestnut blight fungus by mild and severe hypoviruses. J. Virol. 78, 4145-4155 https://doi.org/10.1128/JVI.78.8.4145-4155.2004
  3. Anagnostakis, S.L. 1982. Biological control of chestnut blight. Science 215, 466-47 https://doi.org/10.1126/science.215.4532.466
  4. Chen, B., S. Gao, G.H. Choi, and D.L. Nuss. 1996. Extensive alteration of fungal gene transcript accumulation and elevation of G-protein-regulated cAMP levels by a virulence-attenuating hypovirus. Proc. Natl. Acad. Sci. USA 93, 7996-8000
  5. Cho, Y.W., Y.H. Shin, Y.T. Kim, H.G. Kim, Y.J. Lee, E.H. Park, J.A. Fuchs, and C.J. Lim. 2001. Characterization and regulation of Schizosaccharomyces pombe gene encoding thioredoxin. Biochim. Biophys. Acta. 1518, 194-199 https://doi.org/10.1016/S0167-4781(01)00176-2
  6. Choi, G.H., T.G. Larson, and D.L. Nuss. 1992. Molecular analysis of the laccase gene from the chestnut blight fungus and selective expression of its expression in an isogenic hypovirulent strain. Mol. Plant Microbe Interact. 5, 119-128 https://doi.org/10.1094/MPMI-5-119
  7. Elliston, J.E. 1985. Characteristics of dsRNA-free and dsRNA-containing strains of Endothia parasitica in relation to hypovirulence. Phytopathology 75, 151-158 https://doi.org/10.1094/Phyto-75-151
  8. Eklund, H., F.K. Gleason, and A. Holmgren. 1991. Structural and functional relations among thioredoxins of different species. Proteins 11, 13-28 https://doi.org/10.1002/prot.340110103
  9. Gao, S. and D.L. Nuss. 1996. Distinct roles for two G protein alpha subunits in fungal virulence, morphology, and reproduction revealed by targeted gene disruption. Proc. Natl. Acad. Sci. USA 93, 14122-14127
  10. Gurr, S.J., S.E. Unkles, and J.R. Kinghorn. 1987. The structure and organization of nuclear gene in filamentous fungi, p. 93-139. In J.R. Kinghorn (ed.), Gene structure in eukaryotic microbes, IRL Press, Oxford, UK
  11. Hassan, H.M. and I. Fridovich. 1979. Intracellular production of super-oxide radical and of hydrogen peroxide by redox active compounds. Arch. Biochem. Biophys. 196, 385-395 https://doi.org/10.1016/0003-9861(79)90289-3
  12. Havir, E.A. and S.L. Anagnostakis. 1983. Oxalate production by virulent but not by hypovirulent strains of Endothia parasitica. Physiol. Plant Pathol. 23, 369-376 https://doi.org/10.1016/0048-4059(83)90021-8
  13. Ju, YJ., K.H. Park, D.H. Kim, and B.J. Cha. 1999. Survey on chestnut blight caused by Cryphonectria parasitica. Program Abstr. Annual Meeting of the Korean Society of Plant Pathology. Abstr. p. 13
  14. Kang, H.S., J.W. Choi, S.M. Park, B.J. Cha, M.S. Yang, and D.H. Kim. 1999. Ordered differential display from Cryphonectria parasitica. J. Plant Pathology 16, 142-146
  15. Kasahara, S. and D.L. Nuss. 1997. Targeted disruption of a fungal G-protein beta subunit gene results in increased vegetative growth but reduced virulence. Mol. Plant Microbe Interact. 10, 984-993 https://doi.org/10.1094/MPMI.1997.10.8.984
  16. Kazmierczak, P., P. Pfeiffer, L. Zhang, and N.K. Van Alfen. 1996. Transcriptional repression of specific host genes by the mycovirus Cryphonectria hypovirus CHVI. J. Virol. 70, 1137-1142
  17. Kim, D.H., D. Rigling, L. Zhang, and N.K. Van Alfen, 1995. A new extracellular laccase of Cryphonectria parasitica is revealed by deletion of LacA. Mol. Plant Microbe Interact. 8, 259-266 https://doi.org/10.1094/MPMI-8-0259
  18. Kim, M.J., J.W. Choi, S.M. Park, B.J. Cha, M.S. Yang, and D.H. Kim. 2002. Characterization of a fungal protein kinase from Cryphonectria parasitica and its transcriptional upregulation by hypovirus. Mol. Microbiol. 45, 933-941 https://doi.org/10.1046/j.1365-2958.2002.03079.x
  19. Laurent, T.C., E.C. Moore, and P. Reichard. 1964. Enzymatic synthesis of deoxyribonucleotides IV. Isolation and characterization of thioredoxin the hydrogen donor from E. coli. J. Biol. Chem. 239, 3436-3444
  20. Lee, J.K., H.Y. Lee, S.H. Lee, and M.S. Hwang. 1999. Assessment of tree susceptibility to chestnut blight fungus (Cryphonectria parasitica) of various chestnut varieties. Program Abstr. Annual Meeting of the Korean Society of Plant Pathology. Abstr. p. 45
  21. Luthman, M. and A. Holmgren. 1982. Rat liver thioredoxin and thioredoxin reductase: purification and characterization. Biochemistry 21, 6628-6633 https://doi.org/10.1021/bi00269a003
  22. Nuss, D.L. 1992. Biological control of chestnut blight: an example of virus-mediated attenuation of fungal pathogenesis. Microbiol. 56, 561-576
  23. Park, S.M., E.S. Choi, M.J. Kim, B.J. Cha, M.S. Yang, and D.H. Kim. 2004. Characterization of HOG1 homologue, CpMK1, from Cryphonectria parasitica and evidence for hypovirus-mediated perturbation of its phosphorylation in response to hypertonic stress. Mol. Microbiol. 51, 1267-1277 https://doi.org/10.1111/j.1365-2958.2004.03919.x
  24. Puhalla, J.E. and S.L. Anagnostakis. 1971. Genetics and nutritional requirements of Endothia parasitca. Phytopathology 61, 169-173 https://doi.org/10.1094/Phyto-61-169
  25. Rigling, D., U. Heiniger, and H.R. Hohl. 1989. Reduction of laccase activity in dsRNA-containing hypovirulent strains of Cryphonectria (Endothia) parasitica. Phytopathology 79, 219-223 https://doi.org/10.1094/Phyto-79-219
  26. Roberts, A.N., V. Berlin, K.M. Hager, and C. Yanofsky. 1988. Molecular analysis of a Neurospora crassa gene expressed during conidiation. Mol. Cell BioI. 8, 2411-2418 https://doi.org/10.1128/MCB.8.6.2411
  27. Sambrook, J., E.F. Fritsch, and T. Maniatis. 1989. Molecular cloning : a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA
  28. Segers, G.C. and D.L. Nuss. 2003. Constitutively activated G negatively regulates virulence, reproduction and hydrophobin gene expression in the chestnut blight fungus Cryphonectria parasitica. Fung. Genet. Biol. 38, 198-208 https://doi.org/10.1016/S1087-1845(02)00534-0
  29. Trotter, E.W. and C.M. Grant. 2002. Thioredoxins are required for protection against a reductive stress in the yeast Saccharomyces cerevisiae. Mol. Microbiol. 46, 869-878 https://doi.org/10.1046/j.1365-2958.2002.03216.x
  30. Van Alfen, N.K., R.A. Jaynes, S.L. Anagnostakis, and P.R. Day. 1975. Chestnut blight: biological control by transmissible hypovirulence In Endothia parasitica. Science 189. 890-891 https://doi.org/10.1126/science.189.4206.890
  31. Van Alfen, N.K. 1982. Biology and potential for disease control of hypovirulence of Endothia parasitica. Ann. Rev. Phytopathol. 20, 349-362 https://doi.org/10.1146/annurev.py.20.090182.002025
  32. Zhang, L., D. Villalon, Y. Sun, P. Kazmierczak, and N.K. Van Alfen. 1994. Virus-associated down-regulation of the gene encoding cryparin, an abundant cell-surface protein from the chestnut blight fungus, Cryphonectria parasitica. Gene 139. 59-64