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Volume 140, Issue 11

Partial characterization of specific inducers of a cuticle-degrading protease from the insect pathogenic fungus Free

Abstract

The insect pathogenic fungus produces several extracellular cuticle-degrading proteases and evidence is consistent with one of these, PR1, which is a chymoelastase, being a determinant of pathogenicity. We have shown previously that PR1 production is regulated by both carbon catabolite and nitrogen metabolite repression and also by specific induction under derepressed conditions by insect cuticle. In the present work we have established that an enzymically released proteinaceous component(s) of insect cuticle is capable of inducing PR1 (based on appearance of extracellular activity). Cuticle of the desert locust treated with KOH to remove protein failed to induce PR1 production, whereas cuticle treated with either chloroform or ether to remove lipids still induced PR1. Cuticle digested with either PR1 or the trypsin-like PR2 of released peptides mainly in the range 150-2000 Da; addition of these peptides generated by PR1 or PR2 at 3 μg alanine equivalents mlinduced PR1 production to a level similar (75%) to that obtained with untreated insect cuticle. Several amino acids and peptides which are abundant in insect cuticular protein (Ala, Gly, Ala-Ala, Ala-Ala-Ala, Ala-Pro and Pro-Ala) were tested at a range of concentrations and in restricted cultures for their ability to induce PR1. None induced the protease to the levels seen with cuticle or peptides enzymically released from cuticle, although some dimers and notably the monomers Ala and Gly gave 2-2-7-fold enhanced PR1 activity above derepressed basal levels (up to 48-57% of that achieved with induced synthesis on cuticle). There was evidence for more efficient uptake and/or catabolism by of alanine di-and tripeptides than of monomer amino acids.

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Keyword(s): insect pathogen , Metarhizium anisopliae and protease regulation
© Society for General Microbiology 1994
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1994-11-01
2024-04-30
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References

  1. Andersen S.O. 1979; Biochemistry of insect cuticle.. Annu Rep Entornol 24:29–61
     [Google Scholar]
  2. Andersen S.O. 1988; Comparison between cuticular proteins from two locust species, Locusta migratoria and Schistocerca gregaria.. Insect Biochem 18:751–760
     [Google Scholar]
  3. Andersen J. S., Andersen S.O., Hojrup P., Roepstrorff P. 1993; Primary structure of a 14 kDa basic structural protein (Lm- 76) from the cuticle of the migratory locust Locusta migratoria.. Invertebr Biochem Mol Biol 23:391–402
     [Google Scholar]
  4. Charnley A.K. 1984; Physiological aspects of destructive pathogenesis in insects by fungi: a speculative review. In Invertebrate Microbial Interactions. British Mycological Society Symposium pp. 229–270 Edited by Anderson J. M., Rayner M. A. D., Walton H.D. W. 6: London: Cambridge University Press;
     [Google Scholar]
  5. Charnley A.K., St Leger R.J. 1991; The role of cuticle- degrading enzymes in fungal pathogenesis of insects.. In The Fungal Spore in Disease Initiation in Liants and Animals pp. 267–287 Edited by Cole G. T., Hoch H. C. . New York & London: Plenum Press;
     [Google Scholar]
  6. Cole S.C.J., Charnley A. K., Cooper R. M. 1993; Purification and partial characterisation of a novel trypsin-like cysteine protease from Metarhinfium anisopliae.. FEMS Microbiol Lett 113:189–196
     [Google Scholar]
  7. Cooper R.M. 1977; Regulation of synthesis of cell wall-degrading enzymes of plant pathogens.. In Cell Wall Biochemistry Related to Specificity in Host Plant Interactions pp. 163–211 Edited by Solheim B., Raa J. . Oslo: Universitetsforlaget;
     [Google Scholar]
  8. Cooper R.M., Wood S.R. K. 1975; Regulation of synthesis of cell wall-degrading enzymes by Verticillium albo-atrum and Fusarium oxysporum f. sp. lycopersici.. Physiol Plant Pathol 5:135–156
     [Google Scholar]
  9. Gilby A.R. 1980; Chemical methods lipids. In Cuticle Techniques in Arthropods pp. 217–252 Edited by Miller T. A. Springer-Verlag: New York;
     [Google Scholar]
  10. Grosscurt A.C. 1978; Effects of difluorbenzuron on mechanical penetrability, chitin formation and structure of the elytra of Leptinotarsa decemlineata.. J Insect Physiol 24:827–831
     [Google Scholar]
  11. Gupta D.P., Heale J. B. 1971; Induction of cellulase (Cx) in Verticillium albo-atrum.. J Gen Microbiol 63:163–173
     [Google Scholar]
  12. Henzel W. J., Mole J. E., Mulligan K., Lipke H. 1985; Sarcophagid larval proteins: partial sequence homologies among three cuticle proteins and related structures of Drosophilids.. J Mol Evol 22:39–45
     [Google Scholar]
  13. Hojrup P., Andersen S. O., Roepstrorff P. 1986a; Isolation, characterisation and N-terminal sequence studies of cuticular proteins from the migratory locust, Locusta migratoria.. Eur J Biochem 154:153–159
     [Google Scholar]
  14. Hojrup P., Andersen S.O., Roepstrorff P. 1986b; Primary structure of a structural protein from the cuticle of the migratory locust, Locusta migratoria.. Biochem J 236:713–720
     [Google Scholar]
  15. Klarskov K., Hojrup P., Andersen S. O., Roepstrorff P. 1989; Plasma-desorption mass spectrometry as an aid in protein sequence determination — application of the method on a cuticular protein from the migratory locust, Locusta migratoria.. Biochem J 262:923–930
     [Google Scholar]
  16. Lasure L.L. 1980; Regulation of extracellular acid protease in Mucor miehei.. Mycologia 72:483–492
     [Google Scholar]
  17. Lerner C.G., Goldman R. C. 1993; Stimuli that induce production of Candida albicans extracellular aspartyl proteinase.. J Gen Microbiol 139:1643–1651
     [Google Scholar]
  18. Moore S., Stein W. H. 1948; Photometric ninhydrin method for use in the chromatography of amino acids.. J Biol Chem 76:367–388
     [Google Scholar]
  19. Paterson I. C., Charnley A. K., Cooper R. M., Clarkson J. M. 1993; Regulation of production of a trypsin-like protease by the insect pathogenic fungus Metarhiafium anisopliae.. FEMS Microbiol Lett 109:323–328
     [Google Scholar]
  20. Paterson I. C., Charnley A. K., Cooper R. M., Clarkson J. M. 1994; Specific induction of a cuticle-degrading protease of the insect pathogenic fungus Metarhifium anisopliae.. Microbiology 140:185–189
     [Google Scholar]
  21. Pirt S.J. 1971; The diffusion capsule, a novel device for the addition of a solute at a constant rate to a liquid medium.. Biochem J 121:293–297
     [Google Scholar]
  22. Rebers J.E., Riddiford L. M. 1988; Structure and expression of a Manduca sexta larval cuticle gene homologous to Drosophila genes.. J Mol Biol 203:411–423
     [Google Scholar]
  23. Snyder M., Hunkapillar M., Yeun D., Silver D.J., Fristrom J. W., Davidson N. 1982; Cuticle protein genes of Drosophila: structure, organisation and evolution of four clustered genes.. Cell 29:1027–1040
     [Google Scholar]
  24. St Leger R. J., Cooper R. M., Charnley A. K. 1986; Cuticle-degrading enzymes of entomopathogenic fungi : cuticle degradation in vitro by enzymes from entomopathogens.. J Invertebr Pathol 47:167–177
     [Google Scholar]
  25. St Leger R. J., Charnley A. K., Cooper R. M. 1987a; Characterisation of cuticle-degrading proteases produced by the entomopathogen Metarhizium anisopliae.. Arch Biochem Biophys 253:221–232
     [Google Scholar]
  26. St Leger R. J., Cooper R. M., Charnley A. K. 1987b; Production of cuticle-degrading enzymes by the entomopathogen Metarhizium anisopliae during infection of cuticles from Calliphora vomitoria and Manduca sexta.. J Gen Microbiol 133:1371–1382
     [Google Scholar]
  27. St Leger R. J., Cooper R. M., Charnley A. K. 1987c; Distribution of chymoelastases and trypsin-like enzymes in five species of entomopathogenic Deuteromycetes.. Arch Biochem Biophys 258:123–131
     [Google Scholar]
  28. St Leger R. J., Durrands P. K., Charnley A. K., Cooper R. M. 1988a; Role of extracellular chymoelastase on the virulence of Metarhizium anisopliae for Manduca sexta.. J Invertebr Pathol 52:285–293
     [Google Scholar]
  29. St Leger R. J., Durrands P. K., Cooper R. M., Charnley A. K. 1988b; Regulation of production of proteolytic enzymes by the entomopathogenic fungus Metarhizium anisopliae.. Arch Microbiol 150:413–416
     [Google Scholar]
  30. Talbo G., Hojrup P., Rahbek-Nielson H., Andersen S. O., Roepstrorff P. 1991; Determination of the covalent structure of an N- and C-terminally blocked glycoprotein from the endocuticle of Locusta migratoria.. Eur J Biochem 195:495–504
     [Google Scholar]
  31. Woloshuk C.P., Kolattukudy P. E. 1986; Mechanism by which contact with plant cuticle triggers cutinase gene expression in the spores of Fusarium solani f.sp. pisi.. Proc Natl Acad Sci USA 83:1704–1708
     [Google Scholar]
  32. Weldon C., Kefatos F. C. 1980; Structure, organisation and evolution of developmentally regulated chorion genes in a silk- moth.. Cell 22:855–867
     [Google Scholar]
  33. Willis J.H. 1987; Cuticular proteins: the neglected component.. Arch Insect Biochem Physiol 6:203–215
     [Google Scholar]
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Partial characterization of specific inducers of a cuticle-degrading protease from the insect pathogenic fungus Metarhizium anisopliae
Microbiology 140, 3153 (1994); https://doi.org/10.1099/13500872-140-11-3153
/content/journal/micro/10.1099/13500872-140-11-3153
/content/journal/micro/10.1099/13500872-140-11-3153
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