1887

Abstract

Chitin synthase in a microsomal preparation from had an apparent for UDP--acetylglucosamine of 2.0 mM while nikkomycin Z and polyoxin D inhibited enzyme activity competitively with apparent values of approximately 0.1 μM and 6 μM respectively. The organophosphorus fungicide edifenphos was a non-competitive inhibitor ( 54 μM). Preincubation of microsomes for 2 h at 25 °C resulted in a maximum twofold stimulation of chitin synthase activity while preincubation with trypsin (25 μg ml) or cytosol (350 μg cytosolic protein ml) for 10 min at 25 °C resulted in approximately fourfold and 20-fold increases in chitin synthase activity, respectively. A range of protease inhibitors reduced the degree of activation of microsomal chitin synthase by cytosol. Most potent were phenylmethanesulphonyl fluoride and chymostatin; these compounds completely inhibited activation of enzyme activity. Two fragments (approx. 600 bp; and ) were amplified from genomic DNA using degenerate PCR primers based on regions of complete amino acid homology between previously published chitin synthase gene sequences. When the DNA and predicted amino acid sequences of were used to probe computer databases for related sequences, was found to be most similar to from .

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-9-2199
1994-09-01
2024-12-09
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/9/mic-140-9-2199.html?itemId=/content/journal/micro/10.1099/13500872-140-9-2199&mimeType=html&fmt=ahah

References

  1. Adams D.J. 1994; Drug and pesticide resistance in fungi. In Molecular Genetics of Drug Resistance. Edited by Wolf C. R., Hayes J. D. . London: Harwood Academic Publishers (in press);
    [Google Scholar]
  2. Adams D. J., Causier B. E., Mellor K. J., Keer V., Milling R., Dada J. 1993; Regulation of chitin synthase and chitinase in fungi. In Chitin Enzymology pp. 15–25 Edited by Muzzarelli R.A. A. Ancona: European Chitin Society;
    [Google Scholar]
  3. Backhouse D., Willets H. J. 1984; A histochemical study of sclerotia of Botrjtis cinerea and Botrytis fabae.. Can J Microbiol 30:171–178
    [Google Scholar]
  4. Beynon R.J., Salvesen G. 1989; Commercially available protease inhibitors. In Proteolytic Enzymes. A Practical Approach pp. 241–249 Edited by Beynon R. J., Bond J. S. . Oxford: IRL Press;
    [Google Scholar]
  5. Binks P. R., Robson G. D., Goosey M. W., Humphreys A., Trinci A. P. J. 1991; Chitin synthesis in Fusarium graminearum and its inhibition by edifenphos (Hinosan).. J Gen Microbiol 137:615–620
    [Google Scholar]
  6. 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 Set USA 87:7424–7428
    [Google Scholar]
  7. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  8. Cabib E. 1987; The synthesis and degradation of chitin. Adv Envy mol 59:59–101
    [Google Scholar]
  9. Cabib E., Farkas V. 1971; The control of morphogenesis: an enzymatic mechanism for the initiation of septum formation in yeasi. Proc Natl Acad Set USA 68:2052–2056
    [Google Scholar]
  10. Cabib E., Silverman S. J., Shaw J. A. 1992; Chitinase and chitin synthase 1: counterbalancing activities in cell separation of Saccharomyces cerevisiae. J Gen Microbiol 138:97–102
    [Google Scholar]
  11. Causier B.E. 1993 Chitin synthase of Botrytis cinerea. PhD thesis University of Leeds;
    [Google Scholar]
  12. Chen-Wu J.L., Zwicker 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
    [Google Scholar]
  13. Cohen E., Elster I., Chet I. 1986; Properties and inhibition of Sclerotium rolfsii chitin synthetase.. Pestic Sci 17:175–182
    [Google Scholar]
  14. Dickinson K., Keer V., Hitchcock C. A., Adams D. J. 1989; Chitinase activity from Candida albicans and its inhibition by allosamidin. J Gen Microbiol 135:1417–1421
    [Google Scholar]
  15. Dickinson K., Keer V., Hitchcock C. A., Adams D. J. 1991; Microsomal chitinase activity from Candida albicans. Biochim Biophys Acta 1073:177–182
    [Google Scholar]
  16. Fiedler H.-P., Kurth R., Langharig J., Delzer J., Zahner H. 1982; Nikkomycins: microbial inhibitors of chitin synthase. J Chem Technol Biotechnol 32:271–280
    [Google Scholar]
  17. Flemming S., Cwielong P., Huttermann A. 1982; The cell wall of Fames annosus (Heterobasidion anno sum) as a target for biochemical control. 2. The influence of inhibitors of cell wall synthesis. Fur J Forest Pathol 12:273–280
    [Google Scholar]
  18. Gooday G.W. 1990; Inhibition of chitin metabolism. In Biochemistry of Cell Walls and Membranes in Fungi pp. 61–79 Edited by Kuhn P. I., Trinci A. P. J. , Jung M. J. , Goosey M. W. , Copping L. G. . Berlin: Springer-Verlag;
    [Google Scholar]
  19. Gooday G.W., Trinci A. P. J. 1980; Wall structure and biosynthesis in fungi. In The Eukaryotic Microbial Cell pp. 207–251 Edited by Gooday G. W., Lloyd D., Trinci A. P. J. . Cambridge: Cambridge University Press;
    [Google Scholar]
  20. Groves J. D., Fox R. T. V., Baldwin B. C. 1988; Modes of action of carbendazim and ethyl N-(3,5-dichlorophenyl)carbamate on field isolates of Botrytis cinerea.. Proceedings of the British Crop Protection Conference Pests and Diseases 1988 pp. 397–402
    [Google Scholar]
  21. Higgins D. G., Bleasby A. J., Fuchs R. 1991; Clustal V: improved software for multiple sequence alignment. Comput Appl Biosci 8:189–191
    [Google Scholar]
  22. Hori M., Kakiki K., Suzuki S., Misato T. 1971; Studies on the mode of action of polyoxins. Part III. Relation of polyoxin structure to chitin synthetase inhibition. Agric Biol Chem 35:1280–1291
    [Google Scholar]
  23. Hori M., Kakiki K., Misato T. 1974a; Further study on the relation of polyoxin structure to chitin synthetase inhibition. Agric Biol Chem 38:691–698
    [Google Scholar]
  24. Hori M., Kakiki K., Misato T. 1974b; Interaction between polyoxin and active centre of chitin synthetase. Agric Biol Chem 38:699–705
    [Google Scholar]
  25. Hori M., Eguchi J., Kakiki K., Misato T. 1974c; Studies on the mode of action of polyoxins. VI. Effect of polyoxin B on chitin synthesis in polyoxin-sensitive and resistant strains of Alternaria kikuchiana.. J Antibiot 21:260–266
    [Google Scholar]
  26. Houslay M.D., Stanley K. K. 1982 Dynamics of Biological Membranes. Influence on Synthesis, Structure and Function. Chichester: John Wiley and Sons;
    [Google Scholar]
  27. Kodama O., Yamashita K., Akatsuka T. 1980; Edifenphos, inhibitor of phosphatidylcholine biosynthesis in Pyricularia oryzae.. Agric Biol Chem 44:1015–1021
    [Google Scholar]
  28. Leroux P., Clerjeau M. 1985; Resistance of Botrytis cinerea Pers. and Plasmopara viticola (Berk. and Curt.) Berl. and de Toni to fungicides in French vineyards.. Crop Prot 4:137–160
    [Google Scholar]
  29. Locke T., Fletcher J. T. 1988; Incidence of benomyl and iprodione resistance in isolates of Botrytis cinerea in tomato crops in England and Wales in 1986.. Plant Pathol 37:381–384
    [Google Scholar]
  30. Lundblad G., Hederstedt B., Lind J., Steby M. 1974; Chitinase in goat serum: primary purification and characterization. Eur J Biochem 46:367–376
    [Google Scholar]
  31. Machida S., Saito M. 1993; Purification and characterization of membrane-bound chitin synthase. J Biol Chem 268:1702–1707
    [Google Scholar]
  32. Murray M.G., Thompson W. F. 1980; Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
    [Google Scholar]
  33. Pommer E.-H., Lorenz G. 1982; Resistance of Botrytis cinerea Pers. to dicarboximide fungicides a literature review.. Crop Prot 1:221–230
    [Google Scholar]
  34. Rast D. M., Horsch M., Furter R., Gooday G. W. 1991; A complex chitinolytic system in exponentially growing mycelium of Mucor rouxii: properties and function. J Gen Microbiol 137:2797–2810
    [Google Scholar]
  35. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  36. Shaw J. A., Mol P. C., Bowers B., Silverman S. J., Valdivieso M. H., Duran A., Cabib E. 1991; The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle.. J Cell Biol 114:111–123
    [Google Scholar]
  37. Yarden O., Yanofsky C. 1991; Chitin synthase 1 plays a major role in cell wall biogenesis in Neurospora crassa. Genes and Dev 5:2420–2430
    [Google Scholar]
  38. Zhu W.-Y., Gooday G. W. 1992; Effects of nikkomycin and echinocandin on differentiated and undifferentiated mycelia of Botrytis cinerea and Mucor rouxii.. My col Res 96:371–377
    [Google Scholar]
/content/journal/micro/10.1099/13500872-140-9-2199
Loading
/content/journal/micro/10.1099/13500872-140-9-2199
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