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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)
Publication Information
Journal of Microbiology / v.44, no.5, 2006 , pp. 556-561 More about this Journal
Abstract
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.
Keywords
Cryphonectria parasitica; hypovirulence; thioredoxin;
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1 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   DOI
2 Anagnostakis, S.L. 1982. Biological control of chestnut blight. Science 215, 466-47   DOI   ScienceOn
3 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   DOI   ScienceOn
4 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   DOI   ScienceOn
5 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
6 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   DOI   ScienceOn
7 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
8 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   DOI   ScienceOn
9 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
10 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
11 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   DOI
12 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
13 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
14 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   DOI
15 Elliston, J.E. 1985. Characteristics of dsRNA-free and dsRNA-containing strains of Endothia parasitica in relation to hypovirulence. Phytopathology 75, 151-158   DOI
16 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   DOI
17 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
18 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   DOI   ScienceOn
19 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
20 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   DOI   ScienceOn
21 Eklund, H., F.K. Gleason, and A. Holmgren. 1991. Structural and functional relations among thioredoxins of different species. Proteins 11, 13-28   DOI   ScienceOn
22 Luthman, M. and A. Holmgren. 1982. Rat liver thioredoxin and thioredoxin reductase: purification and characterization. Biochemistry 21, 6628-6633   DOI   ScienceOn
23 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   DOI   ScienceOn
24 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
25 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   DOI   ScienceOn
26 Van Alfen, N.K. 1982. Biology and potential for disease control of hypovirulence of Endothia parasitica. Ann. Rev. Phytopathol. 20, 349-362   DOI   ScienceOn
27 Nuss, D.L. 1992. Biological control of chestnut blight: an example of virus-mediated attenuation of fungal pathogenesis. Microbiol. 56, 561-576
28 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
29 Puhalla, J.E. and S.L. Anagnostakis. 1971. Genetics and nutritional requirements of Endothia parasitca. Phytopathology 61, 169-173   DOI
30 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   DOI   ScienceOn
31 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   DOI   ScienceOn
32 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   DOI