1887

Abstract

The functions of two previously identified chitin synthase genes in , and , were analysed. First, a conditional mutant was constructed in which the expression of is under the control of a repressible promoter, the promoter, of . Under repressing conditions, the mutant grew slowly and produced highly branched hyphae, supporting the idea that is involved in normal hyphal growth. The involvement of in conidiation was also demonstrated. Next, double mutants of and were constructed, in which was placed under the control of the promoter and was replaced with the gene of . These double mutants grew more slowly than the single mutant under high-osmolarity conditions. The hyphae of the double mutant appeared to be more disorganized than those of the single mutant. It was also found that ChsD was functionally implicated in conidiation when the expression of was limited. These results indicate the importance of the ChsD function in the absence of expression. The roles of ChsB and ChsD in hyphal growth and in conidiation were supported by the analysis of the spatial expression patterns of and , using of as a reporter gene.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-5-1335
2002-05-01
2019-12-08
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/5/1481335a.html?itemId=/content/journal/micro/10.1099/00221287-148-5-1335&mimeType=html&fmt=ahah

References

  1. Adams, T. H. & Timberlake, W. E. ( 1990; ). Upstream elements repress premature expression of an Aspergillus developmental regulatory gene. Mol Cell Biol 10, 4912-4919.
    [Google Scholar]
  2. Aguirre, J., Adams, T. H. & Timberlake, W. E. ( 1990; ). Spacial control of developmental regulatory genes in Aspergillus nidulans. Exp Mycol 14, 290-293.[CrossRef]
    [Google Scholar]
  3. Aufauvre-Brown, A., Mellado, E., Gow, N. A. R. & Holden, D. W. ( 1997; ). Aspergillus fumigatus chsE: a gene related to CHS3 of Saccharomyces cerevisiae and important for hyphal growth and conidiophore development but not pathogenicity. Fungal Genet Biol 21, 141-152.[CrossRef]
    [Google Scholar]
  4. Ballance, D. J. & Turner, G. ( 1985; ). Development of a high-frequency transforming vector for Aspergillus nidulans. Gene 36, 321-331.[CrossRef]
    [Google Scholar]
  5. Beth Din, A. & Yarden, O. ( 1994; ). The Neurospora crassa chs-2 gene encodes a non-essential chitin synthase. Microbiology 140, 2189-2197.[CrossRef]
    [Google Scholar]
  6. Beth Din, A. & Yarden, O. ( 2000; ). The Neurospora crassa chs3 gene encodes an essential class I chitin synthase. Mycologia 92, 65-73.[CrossRef]
    [Google Scholar]
  7. Beth Din, A., Specht, C. A., Robbins, P. W. & Yarden, O. ( 1996; ). chs-4, a class IV chitin synthase gene from Neurospora crassa. Mol Gen Genet 250, 214-222.
    [Google Scholar]
  8. Borgia, P. T., Iartchouk, N., Riggle, P. J., Winter, K. R., Koltin, Y. & Bulawa, C. E. ( 1996; ). The chsB gene of Aspergillus nidulans is necessary for normal hyphal growth and development. Fungal Genet Biol 20, 193-203.[CrossRef]
    [Google Scholar]
  9. Bowen, A. R., Chen-Wu, J. L., Momany, M., Young, R., Szaniszlo, P. J. & Robbins, P. W. ( 1992; ). Classification of fungal chitin synthases. Proc Natl Acad Sci USA 89, 519-523.[CrossRef]
    [Google Scholar]
  10. Casadaban, M. J., Martinez-Arias, A., Shapira, S. K. & Chou, J. ( 1983; ). Beta-galactosidase gene fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol 100, 293-308.
    [Google Scholar]
  11. Chen-Wu, J., Zwicher, J., Bowen, A. R. & Robbins, P. W. ( 1992; ). Expression of chitin synthase genes during yeast and hyphal growth phases of Candida albicans. Mol Microbiol 6, 497-502.[CrossRef]
    [Google Scholar]
  12. Choi, W.-J., Santos, B., Durán, A. & Cabib, E. ( 1994; ). Are yeast chitin synthases regulated at the transcriptional or the posttranslational level? Mol Cell Biol 14, 7685-7694.
    [Google Scholar]
  13. Culp, D. W., Dodge, C. L., Miao, Y., Li, L., Sag-Ozkal, D. & Borgia, P. T. ( 2000; ). The chsA gene from Aspergillus nidulans is necessary for maximal conidiation. FEMS Microbiol Lett 182, 349-353.[CrossRef]
    [Google Scholar]
  14. Fujiwara, M., Horiuchi, H., Ohta, A. & Takagi, M. ( 1997; ). A novel fungal gene encoding chitin synthase with a myosin motor-like domain. Biochem Biophys Res Commun 236, 75-78.[CrossRef]
    [Google Scholar]
  15. Fujiwara, M., Ichinomiya, M., Motoyama, T., Horiuchi, H., Ohta, A. & Takagi, M. ( 2000; ). Evidence that the Aspergillus nidulans class I and class II chitin synthase genes, chsC and chsA, share critical roles in hyphal wall integrity and conidiophore development. J Biochem 127, 359-366.[CrossRef]
    [Google Scholar]
  16. Gold, S. E. & Kronstad, J. W. ( 1994; ). Disruption of two genes for chitin synthase in the phytopathogenic fungus Ustilago maydis. Mol Microbiol 11, 897-902.[CrossRef]
    [Google Scholar]
  17. Horiuchi, H., Fujiwara, M., Yamashita, S., Ohta, A. & Takagi, M. ( 1999; ). Proliferation of intrahyphal hyphae caused by disruption of csmA which encodes a class V-chitin synthase with a myosin motor-like domain in Aspergillus nidulans. J Bacteriol 181, 3721-3729.
    [Google Scholar]
  18. Kulmburg, P., Judewicz, N., Mathieu, M., Lenouvel, F., Sequeval, D. & Felenbok, B. ( 1992; ). Specific binding sites for the activator protein, ALCR, in the alcA promoter of the ethanol regulon of Aspergillus nidulans. J Biol Chem 267, 21146-21153.
    [Google Scholar]
  19. May, G. (1992). Fungal technology. In Applied Molecular Genetics of Filamentous Fungi, pp. 1–27. Edited by J. R. Kinghorn & G. Turner. London: Chapman & Hall.
  20. McGoldrick, C. A., Gruver, C. & May, G. S. ( 1995; ). myoA of Aspergillus nidulans encodes an essential myosin I required for secretion and polarized growth. J Cell Biol 128, 577-587.[CrossRef]
    [Google Scholar]
  21. Mellado, E., Aufauvre-Brown, A., Gow, N. A. R. & Holden, D. W. ( 1996; ). The Aspergillus fumigatus chsC and chsG genes encode Class III chitin synthases with different functions. Mol Microbiol 20, 667-679.[CrossRef]
    [Google Scholar]
  22. Miller, K. Y., Toennis, T. M., Adams, T. H. & Miller, B. L. ( 1991; ). Isolation and transcriptional characterization of a morphological modifier: the Aspergillus nidulans Stunted (stuA) gene. Mol Gen Genet 227, 285-292.
    [Google Scholar]
  23. Motoyama, T., Kojima, N., Horiuchi, H., Ohta, A. & Takagi, M. ( 1994a; ). Isolation of a chitin synthase gene (chsC) of Aspergillus nidulans. Biosci Biotechnol Biochem 58, 2254-2257.[CrossRef]
    [Google Scholar]
  24. Motoyama, T., Sudoh, M., Horiuchi, H., Ohta, A. & Takagi, M. ( 1994b; ). Isolation and characterization of two chitin synthase genes of Rhizopus oligosporus. Biosci Biotechnol Biochem 58, 1685-1693.[CrossRef]
    [Google Scholar]
  25. Motoyama, T., Fujiwara, M., Kojima, N., Horiuchi, H., Ohta, A. & Takagi, M. ( 1996; ). The Aspergillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation. Mol Gen Genet 251, 442–450. (Corrigendum in Mol Gen Genet 253, 520–528.).
    [Google Scholar]
  26. Munro, C. A., Schofield, D. A., Gooday, G. W. & Gow, N. A. R. ( 1998; ). Regulation of chitin synthesis during dimorphic growth of Candida albicans. Microbiology 144, 391-401.[CrossRef]
    [Google Scholar]
  27. Munro, C. A., Winter, K., Buchan, A., Henry, K., Becker, J. M., Brown, A. J., Bulawa, C. E. & Gow, N. A. R. ( 2001; ). Chs1 of Candida albicans is an essential chitin synthase required for synthesis of the septum and for cell integrity. Mol Microbiol 39, 1414-1426.
    [Google Scholar]
  28. Oakley, C. E., Weil, C. F., Kretz, P. L. & Oakley, B. R. ( 1987; ). Cloning of the riboB locus of Aspergillus nidulans. Gene 53, 293-298.[CrossRef]
    [Google Scholar]
  29. Pammer, M., Briza, P., Ellinger, A., Schuster, T., Stucka, R., Feldmann, H. & Breitenbach, M. ( 1992; ). DIT101 (CSD2, CAL1), a cell cycle-regulated yeast gene required for synthesis of chitin in cell walls and chitosan in spore walls. Yeast 8, 1089-1099.[CrossRef]
    [Google Scholar]
  30. Rowlands, R. T. & Turner, G. ( 1973; ). Nuclear and extranuclear inheritance of oligomycin resistance in Aspergillus nidulans. Mol Gen Genet 126, 201-216.[CrossRef]
    [Google Scholar]
  31. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  32. Shaw, J. A., Mol, P. C., Bowers, B., Silverman, S. J., Valdivieso, M. H., Durán, A. & Cabib, E. ( 1991; ). The function of chitin synthase 2 and 3 in the Saccharomyces cerevisiae cell cycle. J Cell Biol 114, 111-123.[CrossRef]
    [Google Scholar]
  33. Som, T. & Kolaparthi, V. S. R. ( 1994; ). Developmental decisions in Aspergillus nidulans are modulated by Ras activity. Mol Cell Biol 14, 5333-5348.
    [Google Scholar]
  34. Specht, C. A., Liu, Y., Robbins, P. W. & 8 other authors ( 1996; ). The chsD and chsE genes of Aspergillus nidulans and their roles in chitin synthesis. Fungal Genet Biol 20, 153–167.[CrossRef]
    [Google Scholar]
  35. Sudoh, M., Nagahashi, S., Doi, M., Ohta, A., Takagi, M. & Arisawa, M. ( 1993; ). Cloning of the chitin synthase 3 gene from Candida albicans and its expression during yeast–hyphal transition. Mol Gen Genet 241, 351-358.
    [Google Scholar]
  36. Sudoh, M., Yamazaki, T., Masubuchi, K., Taniguchi, M., Shimma, N., Arisawa, M. & Yamada-Okabe, H. ( 2000; ). Identification of a novel inhibitor specific to the fungal chitin synthase. Inhibition of chitin synthase 1 arrests the cell growth, but inhibition of chitin synthase 1 and 2 is lethal in the pathogenic fungus Candida albicans. J Biol Chem 275, 32901-32905.[CrossRef]
    [Google Scholar]
  37. Valdivieso, M. H., Mol, P. C., Shaw, J. A., Cabib, E. & Durán, A. ( 1991; ). CAL1, a gene required for activity of chitin synthase 3 in Saccharomyces cerevisiae. J Cell Biol 114, 101-109.[CrossRef]
    [Google Scholar]
  38. Wang, Z. & Szaniszlo, P. J. ( 2000; ). WdCHS3, a gene that encodes a class III chitin synthase in Wangiella (Exophiala) dermatitidis, is expressed differentially under stress conditions. J Bacteriol 182, 874-881.[CrossRef]
    [Google Scholar]
  39. Xoconostle-Cázares, B., León-Ramirez, C. & Ruiz-Herrera, J. ( 1996; ). Two chitin synthase genes from Ustilago maydis. Microbiology 142, 377-387.[CrossRef]
    [Google Scholar]
  40. Xoconostle-Cázares, B., Specht, C. A., Robbins, P. W., Liu, Y., León, C. & Ruiz-Herrera, J. ( 1997; ). Umchs5, a gene coding for a class IV chitin synthase in Ustilago maydis. Fungal Genet Biol 22, 199-208.[CrossRef]
    [Google Scholar]
  41. Yanai, K., Kojima, N., Takaya, N., Horiuchi, H., Ohta, A. & Takagi, M. ( 1994; ). Isolation and characterization of two chitin synthase genes from Aspergillus nidulans. Biosci Biotechnol Biochem 58, 1828-1835.[CrossRef]
    [Google Scholar]
  42. Yarden, O. & Yanofsky, C. ( 1991; ). Chitin synthase 1 plays a major role in cell wall biogenesis in Neurospora crassa. Genes Dev 5, 2420-2430.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-5-1335
Loading
/content/journal/micro/10.1099/00221287-148-5-1335
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error