The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

Identification of Genes Encoding Heat Shock Protein 40 Family and the Functional Characterization of Two Hsp40s, MHF16 and MHF21, in Magnaporthe oryzae

  • Yi, Mi-Hwa (Department of Agricultural Biotechnology, Center for Fungal Genetic Resources and Center for Agricultural Biomaterials, Seoul National University) ;
  • Lee, Yong-Hwan (Department of Agricultural Biotechnology, Center for Fungal Genetic Resources and Center for Agricultural Biomaterials, Seoul National University)
  • Published : 2008.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Magnaporthe oryzae, the causal agent of the rice blast disease, poses a worldwide threat to stable rice production. The large-scale functional characterization of genes controlling the pathogenicity of M. oryzae is currently under way, but little is known about heat shock protein 40 (Hsp40) function in the rice blast fungus or any other filamentous plant pathogen. We identified 25 genes encoding putative Hsp40s in the genome of M. oryzae using a bioinformatic approach, which we designated M. oryzae heat shock protein forty (MHF 1-25). To elucidate the roles of these genes, we characterized the functions of MHF16 and MHF21, which encode type ill and type n Hsp40 proteins, respectively. MHF16 and MHF21 expression was not significantly induced by heat shock, but it was down-regulated by cold shock. Knockout mutants of these genes $({\Delta}$mhf16 and ${\Delta}$mhf21) were viable, but conidiation was severely reduced. Moreover, sectoring was observed in the ${\Delta}mhf16$ mutant when it was grown on oatmeal agar medium. Conidial germination, appressorium formation, and pathogenicity in rice were not significantly affected in the mutants. The defects in conidiation and colony morphology were fully complemented by reintroduction of wild type MHF16 and MHF21 alleles, respectively. These data indicate that MHF16 and MHF21 play important roles in conidiation in the rice blast fungus.

Keywords

References

  1. Ahn, N., Kim, S., Choi, W., Im, K. H. and Lee, Y. H. 2004. Extracellular matrix protein gene, EMP1, is required for appressorium formation and pathogenicity of the rice blast fungus, Magnaporthe grisea. Mol. Cells 17:166-173.
  2. Bonato, M. C., Silva, A. M., Gomes, S. L., Maia, J. C. and Juliani, M. H. 1987. Differential expression of heat-shock proteins and spontaneous synthesis of HSP70 during the life cycle of Blastocladiella emersonii. Eur. J. Biochem. 163:211-220. https://doi.org/10.1111/j.1432-1033.1987.tb10757.x
  3. Burnie, J. P., Carter, T. L., Hodgetts, S. J. and Matthews, R. C. 2006. Fungal heat-shock proteins in human disease. FEMS Microbiol. Rev. 30:53-88. https://doi.org/10.1111/j.1574-6976.2005.00001.x
  4. Caplan, A. J. and Douglas, M. G. 1991. Characterization of YDJ1: a yeast homologue of the bacterial dnaJ protein. J. Cell Biol. 114:609-621. https://doi.org/10.1083/jcb.114.4.609
  5. Cheetham, M. E. and Caplan, A. J. 1998. Structure, function and evolution of DnaJ: conservation and adaptation of chaperone function. Cell Stress Chaperones 3:28-36. https://doi.org/10.1379/1466-1268(1998)003<0028:SFAEOD>2.3.CO;2
  6. Chumley, F.G. and Valent, B. 1990. Genetic analysis of melanindeficient, nonpathogenic mutants of Magnaporthe grisea. Mol. Plant-Microbe. Interact. 3:135-143. https://doi.org/10.1094/MPMI-3-135
  7. Craig, E. A. 1985. The heat shock response. CRC Crit. Rev. Biochem. 18:239-280. https://doi.org/10.3109/10409238509085135
  8. Dean, R. A., Talbot, N. J., Ebbole, D. J., Farman, M. L., Mitchell, T. K., Orbach, M. J., Thon, M., Kulkarni, R., Xu, J. R., Pan, H., Read, N. D., Lee, Y. H., Carbone, I., Brown, D., Oh, Y. Y., Donofrio, N., Jeong, J. S., Soanes, D. M., Djonovic, S., Kolomiets, E., Rehmeyer, C., Li, W., Harding, M., Kim, S., Lebrun, M. H., Bohnert, H., Coughlan, S., Butler, J., Calvo, S., Ma, L. J., Nicol, R., Purcell, S., Nusbaum, C., Galagan, J. E. and Birren, B. W. 2005. The genome sequence of the rice blast fungus Magnaporthe grisea. Nature 434:980-986. https://doi.org/10.1038/nature03449
  9. Ebbole, D. J. 2007. Magnaporthe as a model for understanding host-pathogen interactions. Annu. Rev. Phytopathol. 45:437-456. https://doi.org/10.1146/annurev.phyto.45.062806.094346
  10. Fan, C. Y., Lee, S., Ren, H. Y. and Cyr, D. M. 2004. Exchangeable chaperone modules contribute to specification of type I and type II Hsp40 cellular function. Mol. Biol. Cell. 15:761-773. https://doi.org/10.1091/mbc.E03-03-0146
  11. Gething, M. J. 1997. Protein folding. The difference with prokaryotes. Nature 388:329-331. https://doi.org/10.1038/40979
  12. Gething, M. J. and Sambrook, J. 1992. Protein folding in the cell. Nature 355:33-45. https://doi.org/10.1038/355033a0
  13. 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. https://doi.org/10.1073/pnas.95.11.6108
  14. Heikkila, J. J. 1993. Heat shock gene expression and development. I. An overview of fungal, plant, and poikilothermic animal developmental systems. Dev. Genet. 14:1-5. https://doi.org/10.1002/dvg.1020140102
  15. Jeon, J., Park, S. Y., Chi, M. H., Choi, J., Park, J., Rho, H. S., Kim, S., Goh, J., Yoo, S., Park, J. Y., Yi, M., Yang, S., Kwon, M. J., Han, S. S., Kim, B. R., Khang, C. H., Park, B., Lim, S. E., Jung, K., Kong, S., Karunakaran, M., Oh, H. S., Kim, H., Kang, S., Choi, W. B. and Lee, Y. H. 2007. Genome-wide functional analysis of pathogenicity genes in the rice blast fungus. Nat. Genet. 39:561-565. https://doi.org/10.1038/ng2002
  16. Kaufmann, S. H. 1990. Heat shock proteins and the immune response. Immunol. Today 11:129-136. https://doi.org/10.1016/0920-5861(91)87013-D
  17. Kelly, William L. 1998. The J-domain family and the recruitment of chaperone power. Trends Biochem. Sci. 23:222-227. https://doi.org/10.1016/S0968-0004(98)01215-8
  18. Khang, C. H., Park, S. Y., Lee, Y. H. and Kang, S. 2005. A dual selection based, targeted gene replacement tool for Magnaporthe grisea and Fusarium oxysporum. Fungal Genet. Biol. 42:483-492. https://doi.org/10.1016/j.fgb.2005.03.004
  19. Kim, J. E., Kim, J. C., Jin, J., Yun, S. H. and Lee, Y. W. 2008. Functional characterization of genes located at the aurofusarin biosynthesis gene cluster in Gibberella zeae. Plant Pathol. J. 24:8-16. https://doi.org/10.5423/PPJ.2008.24.1.008
  20. Kim, S., Ahn, I. P., Rho, H. S. and Lee, Y. H. 2005. MHP1, a Magnaporthe grisea hydrophobin gene, is required for fungal development and plant colonization. Mol. Microbiol. 57:1224-1237. https://doi.org/10.1111/j.1365-2958.2005.04750.x
  21. Luke, M. M., Sutton, A. and Arndt, K. T. 1991. Characterization of SIS1, a Saccharomyces cerevisiae homologue of bacterial dnaJ proteins. J. Cell Biol. 114:623-638. https://doi.org/10.1083/jcb.114.4.623
  22. Martineau, C. N., Beckerich, J. M. and Kabani, M. 2007. Flo11pindependent control of "mat" formation by hsp70 molecular chaperones and nucleotide exchange factors in yeast. Genetics 177:1679-1689. https://doi.org/10.1534/genetics.107.081141
  23. Meacham, G. C., Browne, B. L., Zhang, W., Kellermayer, R., Bedwell, D. M. and Cyr, D. M. 1999. Mutations in the yeast Hsp40 chaperone protein Ydj1 cause defects in Axl1 biogenesis and pro-a-factor processing. J. Biol. Chem. 274:34396-34402. https://doi.org/10.1074/jbc.274.48.34396
  24. Mulder, N. J., Apweiler, R., Attwood, T. K., Bairoch, A., Bateman, A., Binns, D., Bradley, P., Bork, P., Bucher, P., Cerutti, L., Copley, R., Courcelle, E., Das, U., Durbin, R., Fleischmann, W., Gough, J., Haft, D., Harte, N., Hulo, N., Kahn, D., Kanapin, A., Krestyaninova, M., Lonsdale, D., Lopez, R., Letunic, I., Madera, M., Maslen, J., McDowall, J., Mitchell, A., Nikolskaya, A. N., Orchard, S., Pagni, M., Ponting, C. P., Quevillon, E., Selengut, J., Sigrist, C. J., Silventoinen, V., Studholme, D. J., Vaughan, R. and Wu, C. H. 2005. InterPro, progress and status in 2005. Nucleic Acids Res. 33:D201-205. https://doi.org/10.1093/nar/gki158
  25. Nishikawa, S. and Endo, T. 1997. The yeast JEM1p is a DnaJ-like protein of the endoplasmic reticulum membrane required for nuclear fusion. J. Biol. Chem. 272:12889-12892. https://doi.org/10.1074/jbc.272.20.12889
  26. Ou, S. H. 1985. Rice Diseases. Kew, England: Commonwealth Mycological Institute.
  27. Park, J., Park, B., Jung, K., Jang, S., Yu, K., Choi, J., Kong, S., Kim, S., Kim, H., Kim, J. F., Blair, J. E., Lee, K., Kang, S. and Lee, Y. H. 2008. CFGP: a web-based, comparative fungal genomics platform. Nucleic Acids Res. 36:D562-571. https://doi.org/10.1093/nar/gkm758
  28. Rensing, L., Monnerjahn, C. and Meyer, U. 1998. Differential stress gene expression during the development of Neurospora crassa and other fungi. FEMS Microbiol. Lett. 168:159-166. https://doi.org/10.1111/j.1574-6968.1998.tb13268.x
  29. Rho, H. S., Kang, S. and Lee, Y. H. 2001. Agrobacterium tumefaciens-mediated transformation of the plant pathogenic fungus, Magnaporthe grisea. Mol. Cells 12:407-411.
  30. Rowley, N., Prip-Buus, C., Westermann, B., Brown, C., Schwarz, E., Barrell, B. and Neupert, W. 1994. Mdj1p, a novel chaperone of the DnaJ family, is involved in mitochondrial biogenesis and protein folding. Cell 77:249-259. https://doi.org/10.1016/0092-8674(94)90317-4
  31. Sambrook, J. and Russell, D. W. 2001. Molecular Cloning: a laboratory manual. 3rd ed. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.
  32. Talbot, N. J., Ebbole, D. J. and Hamer, J. E. 1993. Identification and characterization of MPG1, a gene involved in pathogenicity from the rice blast fungus Magnaporthe grisea. Plant Cell 5:1575-1590. https://doi.org/10.1105/tpc.5.11.1575
  33. Thompson, J. D., Higgins, D. G. and Gibson, T. J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22:4673-4680. https://doi.org/10.1093/nar/22.22.4673
  34. Valent, B. 1990. Rice blast as a model system for plant pathology. Phytopathology 80:33-36. https://doi.org/10.1094/Phyto-80-33
  35. 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. https://doi.org/10.1038/sj.embor.7400172
  36. Werner-Washburne, M., Becker, J., Kosic-Smithers, J. and Craig, E. A. 1989. Yeast Hsp70 RNA levels vary in response to the physiological status of the cell. J. Bacteriol. 171:2680-2688. https://doi.org/10.1128/jb.171.5.2680-2688.1989
  37. Westermann, B. and Neupert, W. 1997. Mdj2p, a novel DnaJ homolog in the mitochondrial inner membrane of the yeast. J. Mol. Biol. 272:477-483. https://doi.org/10.1006/jmbi.1997.1267
  38. Wright, C. M., Fewell, S. W., Sullivan, M. L., Pipas, J. M., Watkins, S. C. and Brodsky, J. L. 2007. The Hsp40 molecular chaperone Ydj1p, along with the protein kinase C pathway, affects cell-wall integrity in the yeast Saccharomyces cerevisiae. Genetics 175:1649-1664. https://doi.org/10.1534/genetics.106.066274
  39. Yu, J., Hu, S., Wang, J., Wong, G. K., Li, S., Liu, B., Deng, Y., Dai, L., Zhou, Y., Zhang, X., Cao, M., Liu, J., Sun, J., Tang, J., Chen, Y., Huang, X., Lin, W., Ye, C., Tong, W., Cong, L., Geng, J., Han, Y., Li, L., Li, W., Hu, G., Li, J., Liu, Z., Qi, Q., Li, T., Wang, X., Lu, H., Wu, T., Zhu, M., Ni, P., Han, H., Dong, W., Ren, X., Feng, X., Cui, P., Li, X., Wang, H., Xu, X., Zhai, W., Xu, Z., Zhang, J., He, S., Xu, J., Zhang, K., Zheng, X., Dong, J., Zeng, W., Tao, L., Ye, J., Tan, J., Chen, X., He, J., Liu, D., Tian, W., Tian, C., Xia, H., Bao, Q., Li, G., Gao, H., Cao, T., Zhao, W., Li, P., Chen, W., Zhang, Y., Hu, J., Liu, S., Yang, J., Zhang, G., Xiong, Y., Li, Z., Mao, L., Zhou, C., Zhu, Z., Chen, R., Hao, B., Zheng, W., Chen, S., Guo, W., Tao, M., Zhu, L., Yuan, L. and Yang, H. 2002. A draft sequence of the rice genome (Oryza sativa L. ssp. indica). Science 296:79-92. https://doi.org/10.1126/science.1068037
  40. Zhong, T. and Arndt, K. T. 1993. The yeast SIS1 protein, a DnaJ homolog, is required for the initiation of translation. Cell 73:1175-1186. https://doi.org/10.1016/0092-8674(93)90646-8

Cited by

  1. MoSNF1 regulates sporulation and pathogenicity in the rice blast fungus Magnaporthe oryzae vol.45, pp.8, 2008, https://doi.org/10.1016/j.fgb.2008.05.003
  2. A Novel Pathogenicity Gene Is Required in the Rice Blast Fungus to Suppress the Basal Defenses of the Host vol.5, pp.4, 2009, https://doi.org/10.1371/journal.ppat.1000401