Skip to main content
SpringerLink
Log in

A DnaJ-like homolog from Cryphonectria parasitica is not responsive to hypoviral infection but is important for fungal growth in both wild-type and hypovirulent strains

  • Published:
Molecules and Cells

Abstract

A DnaJ-like gene, Cpdj1, a molecular chaperone and regulator of Hsp70 in Cryphonectria parasitica, was characterized. The protein product of Cpdj1 gene consists of 379 amino acids with a predicted molecular mass of 40.6 kDa and a pI of 7.79. The deduced protein sequence revealed preservation of the conserved hall-mark J-region and exhibited high homolo y to all known DnaJ-like proteins. Disruption of the Cpdj1 gene resulted in slow growth and produced colonies characterized by retarded growth and deep orange color. Accordingly, reduced virulence of the Cpdj1-null mutant was observed. This reduced growth rate was magnified when the Cpdj1-null mutant was cultured under heat-stress conditions. Reduced conidiation was also observed in the Cpdj1-null mutant, indicating that Cpdj1 gene, although not essential for cell viability, is required for appropriate cellular processes including growth and sporulation. Northern analysis showed that Cpdj1 was constitutively expressed, and when the culture was subject to high temperature, a strong induction of the transcript was observed. No significant difference in the expression and induction pattern of Cpdj1 was observed between virus-free EP155/2 and virus-infected hypovirulent UEP1 strains. However, further severe defects in mycelia rowth and conidiation were observed in the hypovirus-infected Cpdj1-null mutant suggesting that the presence of Cpdj1 is required for mycelia growth and sporulation of the hypovirus-infected strain.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Allen, T.D., Dawe, A.L., and Nuss, D.L. (2003). Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonectri parasitic to infection by virulenceat-tenuating hypoviruses. Eukaryot. Cell 2, 1253–1265.

    Article  CAS  PubMed  Google Scholar 

  • Allen, T.D., and Nuss, D.L. (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.

    Article  CAS  PubMed  Google Scholar 

  • Anagnostakis, S.L. (1982). Biological control of chestnut blight. Science 215, 466–471.

    Article  CAS  PubMed  Google Scholar 

  • Atencio, D.P., and Yaffe, M.P. (1992). MAS5, a yeast homolog of DnaJ involved in mitochondrial protein import. Mol. Cell. Biol. 12, 283–291.

    CAS  PubMed  Google Scholar 

  • Caplan, A.J., Cyr, D.M., and Douglas, M.G. (1992). YDJ1p facilitates polypeptide translocation across different intracellular membranes by a conserved mechanism. Cell 71, 1143–1155.

    Article  CAS  PubMed  Google Scholar 

  • Choi, G.H., and Nuss, D.L. (1990). Nucleotide sequence of the glyceraldehyde-3-phosphate dehydrogenase gene from Cryphonectria parasitica. Nucleic Acids Res. 18, 5566.

    Article  CAS  PubMed  Google Scholar 

  • Choi, G.H., Larson, T.G., and Nuss, D.L. (1992). Molecular analysis of the laccase gene from the chestnut blight fungus and selective suppression of its expression in an isogenic hypovirulent strain. Mol. Plant-Microbe Interact. 5, 119–128.

    CAS  PubMed  Google Scholar 

  • Choi, E.S., Chung, H.J., Kim, M.J., Park, S.M., Cha, B.J., Yang, M.S., and Kim, D.H. (2005). Characterization of the ERK homologue CpMK2 from the chestnut blight fungus Cryphonectria parasitica. Microbiology 151, 1349–1358.

    Article  CAS  PubMed  Google Scholar 

  • Chromy, L.R., Pipas, J.M., and Garcea, R.L. (2003). Chaperone-mediated in vitro assembly of Polyomavorus capsids. Proc. Natl. Acad. Sci. USA 100, 10477–10482.

    Article  CAS  PubMed  Google Scholar 

  • Churchill, A.C.L., Ciufetti, L.M., Hansen, D.R., Van Etten, H.D., and Van Alfen, N.K. (1990). Transformation of the fungal pathogen Cryphonectria parasitica with a variety of heterologous plasmids. Curr. Genet. 17, 25–31.

    Article  CAS  Google Scholar 

  • Cullen, D., Leong, S.A., Wilson, L.J., and Henner, D.J. (1987). Transformation of Aspergillus nidulans with the hygromycin-resistance gene, hph. Gene 57, 21–26.

    Article  CAS  PubMed  Google Scholar 

  • Cyr, D.M., Langer, T., and Douglas, M.G. (1994). DnaJ-like proteins: molecular chaperones and specific regulators of Hsp70. Trends Biochem. Sci. 19, 176–181.

    Article  CAS  PubMed  Google Scholar 

  • Deng, F., Allen, T.D., Hillman, B.I., and Nuss, D.L. (2007). Comparative analysis of alterations in host phenotype and transcript accumulation following hypovirus and mycoreovirus infections of the chestnut blight fungus Cryphonectria parasitica. Eukaryot. Cell 6, 1286–1298.

    Article  CAS  PubMed  Google Scholar 

  • Dey, B., Caplan, A.J., and Boschelli, F. (1996). The Ydj1 molecular chaperone facilitates formation of active p60v-src in yeast. Mol. Biol. Cell. 7, 91–100.

    CAS  PubMed  Google Scholar 

  • Elliston, J.E. (1985). Characteristics of dsRNA-free and dsRNA-containing strains of Endothia parasitica in relation to hypovirulence. Phytopathology 75, 151–158.

    Article  Google Scholar 

  • Fu, Q., Li, S., and Yu, D. (2010). Identification of an Arabidopsis Nodulin-related protein in heat stress. Mol. Cells 29, 77–84.

    Article  CAS  PubMed  Google Scholar 

  • Gurr, S.J., Unkles, S.E., and Kinghorn, J.R. (1987). The structure and organization of nuclear gene in filamentous fungi. In Gene Structure in Eukaryotic Microbes, J.R. Kinghorn, ed. (Oxford: ILR Press), pp. 93–139.

    Google Scholar 

  • Greene, M.K., Maskos, K., and Landry, S.J. (1998). Role of the Jdomain in the cooperation of Hsp40 with Hsp70. Proc. Natl. Acad. Sci. USA 95, 6108–6113.

    Article  CAS  PubMed  Google Scholar 

  • Havir, E.A., and Anagnostakis, S.L. (1983). Oxalate production by virulent but not by hypovirulent strains of Endothia parasitica. Physiol. Plant Pathol. 23, 369–376.

    Article  CAS  Google Scholar 

  • Hu, S.M., Liang, P.H., Hsiao, C.D., and Wang, C. (2002). Characterization of the L399P and R447G mutants of hsc70: the decrease in refolding activity is correlated with an increase in the rate of substrate dissociation. Arch. Biochem. Biophys. 407, 135–141.

    Article  CAS  PubMed  Google Scholar 

  • Johnson, J.L., and Craig, E.A. (2001). An essential role for the substrate-binding region of Hsp40s in Saccharomyces cerevisiae. J. Cell Biol. 152, 851–856.

    Article  CAS  PubMed  Google Scholar 

  • Ju, Y.J., Kim, D.H., and Cha, B.J. (2002). Isolation of Cryphonectria parasitica from cankers on chestnut trees in Korea and characteristics of isolates. Plant Pathol. J. 18, 23–29.

    Google Scholar 

  • Kang, H.S., Choi, J.W., Park, S.M., Cha, B.J., Yang, M.S., and Kim, D.H. (2000). Ordered differential display from Cryphonectria parasitica. Plant Pathol. J. 16, 142–146.

    Google Scholar 

  • Kazmierczak, P., Pfeiffer, P., Zhang, L., and Van Alfen, N.K. (1996). Transcriptional repression of specific host genes by the mycovirus Cryphonectria hypovirus CHV1. J. Virol. 70, 1137–1142.

    CAS  PubMed  Google Scholar 

  • Kelly, W.L. (1998). The J-domain family and the recruitment of chaperone power. Trends Biochem. Sci. 23, 222–227.

    Article  Google Scholar 

  • Kim, J.H., and Kim, D.H. (2006). Cloning and characterization of a thioredoxin gene, CpTrx1, from the chestnut blight fungus Cryphonectria parasitica. J. Microbiol. 44, 556–561.

    CAS  PubMed  Google Scholar 

  • Kim, D.H., Rigling, D., Zhang, L., and Van Alfen, N.K. (1995). A new extracellular laccase of Cryphonectria parasitica is revealed by deletion of Lac1. Mol. Plant-Microbe Interact. 8, 259–266.

    Google Scholar 

  • Kim, M.J., Choi, J.W., Park, S.M., Cha, B.J., Yang, M.S., and Kim, D.H. (2002). Characterization of a fungal protein kinase from Cryphonectria parasitica and its transcriptional upregulation by hypovirus. Mol. Microbiol. 45, 933–941.

    Article  CAS  PubMed  Google Scholar 

  • Kim, M.J., Kwon, B.R., Park, S.M., Chung, H.J., Yang, M.S., Churchill, A.C.L., Van Alfen, N.K., and Kim, D.H. (2008). Promoter analysis of the cell surface-abundant and hypoviral-regulated cryparin gene from Cryphonectria parasitica. Mol. Cells 26, 496–502.

    CAS  PubMed  Google Scholar 

  • Lee, J.K., Tattar, T.A., Berman, P.M., and Mount, M.S. (1992). A rapid method for testing the virulence of Cryphonectria parasitica using excised bark and wood of American chestnut. Phytopathology 82, 1454–1456.

    Article  Google Scholar 

  • Lee, D.H., Sherman, M.Y., and Goldberg, A.L. (1996). Involvement of the molecular chaperone Ydj1 in the ubiquitin-dependent degradation of short-lived and abnormal proteins in Saccharomyces cerevisiae. Mol. Cell. Biol. 16, 4773–4781.

    CAS  PubMed  Google Scholar 

  • Lindquist, S., and Craig, E.A. (1988). The heat-shock proteins. Ann. Rev. Genet. 22, 631–677.

    Article  CAS  PubMed  Google Scholar 

  • Lu, Z., and Cyr, D.M. (1998). Protein folding activity of Hsp70 is modified differentially by the hsp40 co-chaperones Sis1 and Ydj1. J. Biol. Chem. 273, 27824–27830.

    Article  CAS  PubMed  Google Scholar 

  • Niewiarowska, J., D’Halluin, J.C., and Belin, M.T. (1992). Adenovirus capsid proteins interact with HSP70 proteins after penetration in human or rodent cells. Exp. Cell Res. 201, 408–416.

    Article  CAS  PubMed  Google Scholar 

  • Nuss, D.L. (1992). Biological control of chestnut blight: an example of virus-mediated attenuation of fungal pathogenesis. Microbiol. Rev. 56, 561–576.

    CAS  PubMed  Google Scholar 

  • Orbach, M.J., Porro, E.B., and Yanofsky, C. (1986). Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker. Mol. Cell. Biol. 6, 2452–2461.

    CAS  PubMed  Google Scholar 

  • Park, S.M., Choi, E.S., Kim, M.J., Cha, B.J., Yang, M.S., and Kim, D.H. (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.

    Article  CAS  PubMed  Google Scholar 

  • Park, S.M., Kim, J.M., Chung, H.J., Lim, J.Y., Kwon, B.R., Lim, J.G., Kim, J.A., Kim, M.J., Cha, B.J., Lee, S.H., et al. (2008). Occurrence of diverse dsRNA in a Korean population of the chestnut blight fungus, Cryphonectria parasitica. Mycol. Res. 112, 1220–1226.

    Article  CAS  PubMed  Google Scholar 

  • Plesofsky, N. (2004). Heat shock proteins and the stress response. In the Mycota III Biochemistry and Molecular Biology, R., Brambl, and C.A., Marzluf, eds. (Berlin: Springer-Verlag), pp. 143–173.

    Google Scholar 

  • Powell, W.A.J., and Van Alfen, N.K. (1987). Two nonhomologus viruses of Cryphonectria (Endothia) parasitica reduce accumulation of specific virulence-associated polypeptides. J. Bacteriol. 169, 5324–5326.

    CAS  PubMed  Google Scholar 

  • Rigling, D., Heiniger, U., and Hohl, H.R. (1989). Reduction of laccase activity in dsRNA-containing hypovirulent strains of Cryphonectria (Endothia) parasitica. Phytopathology 79, 219–223.

    Article  CAS  Google Scholar 

  • Roberts, A.N., Berlin, V., Hager, K.M., and Yanofsky, C. (1988). Molecular analysis of a Neurospora crassa gene expressed during conidiation. Mol. Cell. Biol. 8, 2411–2418.

    CAS  PubMed  Google Scholar 

  • Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular cloning: a laboratory manual, 2nd edition, (NY, USA: Cold Spring Harbor Laboratory Press).

    Google Scholar 

  • Silver, P.A., and Way, J.C. (1993). Eukaryotic DnaJ homologs and the specificity of Hsp70 activity. Cell 74, 5–6.

    Article  CAS  PubMed  Google Scholar 

  • Sullivan, C.S., and Pipas, J.M. (2001). The virus-chaperone connection. Virology 287, 1–8.

    Article  CAS  PubMed  Google Scholar 

  • Sun, Q., Choi, G., and Nuss, D.L. (2009). A silencing argonaute gene is required for induction of RNA silencing antiviral defense and promotes viral RNA recombination. Proc. Natl. Acad. Sci. USA 106, 17927–17932.

    Article  CAS  PubMed  Google Scholar 

  • Tachibana, T., Astumi, S., Shioda, R., Ueno, M., Uritani, M., and Ushimaru, T. (2002). A novel non-conventional heat shock element regulates expression of MDJ1 encoding a DnaJ homolog in Saccharomyces cerevisiae. J. Biol. Chem. 277, 22140–22146.

    Article  CAS  PubMed  Google Scholar 

  • Walsh, P., Bursac, D., Law, Y.C., Cyr, D., and Lithgow, T. (2004). The J-protein family: modulating protein assembly, disassembly and translocation. EMBO Rep. 5, 567–571.

    Article  CAS  PubMed  Google Scholar 

  • Walter S., and Buchner, J. (2002). Molecular chaperones-cellular machines for protein folding. Angew. Chem. Int. Ed. Engl. 41, 1098–1113.

    Article  CAS  PubMed  Google Scholar 

  • Van Alfen, N.K. (1982). Biology and potential for disease control of hypovirulence of Endothia parasitica. Ann. Rev. Phytopathol. 20, 349–362.

    Article  Google Scholar 

  • Van Alfen, N.K., Jaynes, R.A., Anagnostakis, S.L., and Day, P.R. (1975). Chestnut blight: biological control by transmissible hypovirulence in Endothia parasitica. Science 189, 890–891.

    Article  PubMed  Google Scholar 

  • Wang, R.Y., Stork, J., and Nagy, P.D. (2009). A key role for heat shock protein 70 in the localization and insertion of tombusvirus replication proteins to intracellular membranes. J. Virol. 83, 15–22.

    Google Scholar 

  • Zhang, L., Villalon, D., Sun, Y., Kazmierczak, P., and Van Alfen, N.K. (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.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, Chonbuk National University, Jeonju, 561-756, Korea

    Jung-Gu Lim, Joong-Gi Lee, Jung-Mi Kim, Jin-Ah Park, Seung-Moon Park, Moon-Sik Yang & Dae-Hyuk Kim

Authors
  1. Jung-Gu Lim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joong-Gi Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jung-Mi Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jin-Ah Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Seung-Moon Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Moon-Sik Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dae-Hyuk Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dae-Hyuk Kim.

About this article

Cite this article

Lim, JG., Lee, JG., Kim, JM. et al. A DnaJ-like homolog from Cryphonectria parasitica is not responsive to hypoviral infection but is important for fungal growth in both wild-type and hypovirulent strains. Mol Cells 30, 235–243 (2010). https://doi.org/10.1007/s10059-010-0112-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10059-010-0112-1

Keywords

Search

Navigation