Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1996 Aug;111(4):1209–1218. doi: 10.1104/pp.111.4.1209

Mutualistic fungal endophytes express a proteinase that is homologous to proteases suspected to be important in fungal pathogenicity.

P V Reddy 1, C K Lam 1, F C Belanger 1
PMCID: PMC160998  PMID: 8756501

Abstract

Many cultivated and wild grass species are hosts to mutualistic fungal endophytes. These associations are ecologically and agronomically significant, yet little is known regarding the physiological aspects of the interaction. In the Poa ampla/Acremonium typhinum interaction, a fungal serine proteinase, At1, is surprisingly abundant and may constitute 1 to 2% of the total leaf-sheath protein. Sequence analysis of cDNA and genomic clones indicates that proteinase At1 is a member of the eukaryotic subtilisin-like protease family. It is homologous to proteases suspected to be virulence factors in fungal pathogens of insects, nematodes, and other fungi. Gel blot analysis of RNA extracted from infected leaf-sheath tissue indicates that the proteinase At1 transcript level is extremely high. RNA gel blots and immunoblots of purified enzymes indicate that similar proteinases are produced by Epichloë festucae and Acremonium lolii, the fungal endophytes infecting Festuca rubra subsp. rubra and Lolium perenne, respectively. Fungal expression of proteinase At1-like enzymes may be a general feature of endophyte infection.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bacon C. W., Porter J. K., Robbins J. D. Toxicity and occurrence of Balansia on grasses from toxic fescue pastures. Appl Microbiol. 1975 Apr;29(4):553–556. doi: 10.1128/am.29.4.553-556.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Betzel C., Pal G. P., Struck M., Jany K. D., Saenger W. Active-site geometry of proteinase K. Crystallographic study of its complex with a dipeptide chloromethyl ketone inhibitor. FEBS Lett. 1986 Mar 3;197(1-2):105–110. doi: 10.1016/0014-5793(86)80307-6. [DOI] [PubMed] [Google Scholar]
  3. Bonants P. J., Fitters P. F., Thijs H., den Belder E., Waalwijk C., Henfling J. W. A basic serine protease from Paecilomyces lilacinus with biological activity against Meloidogyne hapla eggs. Microbiology. 1995 Apr;141(Pt 4):775–784. doi: 10.1099/13500872-141-4-775. [DOI] [PubMed] [Google Scholar]
  4. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  5. Cheevadhanarak S., Renno D. V., Saunders G., Holt G. Cloning and selective overexpression of an alkaline protease-encoding gene from Aspergillus oryzae. Gene. 1991 Dec 1;108(1):151–155. doi: 10.1016/0378-1119(91)90501-2. [DOI] [PubMed] [Google Scholar]
  6. Chu B., Lindstrom J. T., Belanger F. C. Arabidopsis thaliana expresses three divergent Srp54 genes. Plant Physiol. 1994 Nov;106(3):1157–1162. doi: 10.1104/pp.106.3.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Don R. H., Cox P. T., Wainwright B. J., Baker K., Mattick J. S. 'Touchdown' PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res. 1991 Jul 25;19(14):4008–4008. doi: 10.1093/nar/19.14.4008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fletcher L. R., Harvey I. C. An association of a Lolium endophyte with ryegrass staggers. N Z Vet J. 1981 Oct;29(10):185–186. doi: 10.1080/00480169.1981.34839. [DOI] [PubMed] [Google Scholar]
  9. Frederick G. D., Rombouts P., Buxton F. P. Cloning and characterisation of pepC, a gene encoding a serine protease from Aspergillus niger. Gene. 1993 Mar 15;125(1):57–64. doi: 10.1016/0378-1119(93)90745-o. [DOI] [PubMed] [Google Scholar]
  10. Geremia R. A., Goldman G. H., Jacobs D., Ardiles W., Vila S. B., Van Montagu M., Herrera-Estrella A. Molecular characterization of the proteinase-encoding gene, prb1, related to mycoparasitism by Trichoderma harzianum. Mol Microbiol. 1993 May;8(3):603–613. doi: 10.1111/j.1365-2958.1993.tb01604.x. [DOI] [PubMed] [Google Scholar]
  11. Holden D. W., Tang C. M., Smith J. M. Molecular genetics of Aspergillus pathogenicity. Antonie Van Leeuwenhoek. 1994;65(3):251–255. doi: 10.1007/BF00871953. [DOI] [PubMed] [Google Scholar]
  12. Isogai T., Fukagawa M., Kojo H., Kohsaka M., Aoki H., Imanaka H. Cloning and nucleotide sequences of the complementary and genomic DNAs for the alkaline protease from Acremonium chrysogenum. Agric Biol Chem. 1991 Feb;55(2):471–477. [PubMed] [Google Scholar]
  13. Jarai G., Kirchherr D., Buxton F. P. Cloning and characterization of the pepD gene of Aspergillus niger which codes for a subtilisin-like protease. Gene. 1994 Feb 11;139(1):51–57. doi: 10.1016/0378-1119(94)90522-3. [DOI] [PubMed] [Google Scholar]
  14. Jaton-Ogay K., Paris S., Huerre M., Quadroni M., Falchetto R., Togni G., Latgé J. P., Monod M. Cloning and disruption of the gene encoding an extracellular metalloprotease of Aspergillus fumigatus. Mol Microbiol. 1994 Dec;14(5):917–928. doi: 10.1111/j.1365-2958.1994.tb01327.x. [DOI] [PubMed] [Google Scholar]
  15. Jaton-Ogay K., Suter M., Crameri R., Falchetto R., Fatih A., Monod M. Nucleotide sequence of a genomic and a cDNA clone encoding an extracellular alkaline protease of Aspergillus fumigatus. FEMS Microbiol Lett. 1992 Apr 15;71(2):163–168. doi: 10.1016/0378-1097(92)90506-j. [DOI] [PubMed] [Google Scholar]
  16. Kothary M. H., Chase T., Jr, Macmillan J. D. Correlation of elastase production by some strains of Aspergillus fumigatus with ability to cause pulmonary invasive aspergillosis in mice. Infect Immun. 1984 Jan;43(1):320–325. doi: 10.1128/iai.43.1.320-325.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lindstrom J. T., Belanger F. C. Purification and Characterization of an Endophytic Fungal Proteinase That Is Abundantly Expressed in the Infected Host Grass. Plant Physiol. 1994 Sep;106(1):7–16. doi: 10.1104/pp.106.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marchuk D., Drumm M., Saulino A., Collins F. S. Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res. 1991 Mar 11;19(5):1154–1154. doi: 10.1093/nar/19.5.1154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moehle C. M., Tizard R., Lemmon S. K., Smart J., Jones E. W. Protease B of the lysosomelike vacuole of the yeast Saccharomyces cerevisiae is homologous to the subtilisin family of serine proteases. Mol Cell Biol. 1987 Dec;7(12):4390–4399. doi: 10.1128/mcb.7.12.4390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Samuels R. I., Paterson I. C. Cuticle degrading proteases from insect moulting fluid and culture filtrates of entomopathogenic fungi. Comp Biochem Physiol B Biochem Mol Biol. 1995 Apr;110(4):661–669. doi: 10.1016/0305-0491(94)00205-9. [DOI] [PubMed] [Google Scholar]
  22. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shinde U., Inouye M. Intramolecular chaperones and protein folding. Trends Biochem Sci. 1993 Nov;18(11):442–446. doi: 10.1016/0968-0004(93)90146-e. [DOI] [PubMed] [Google Scholar]
  24. Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. St Leger R. J., Frank D. C., Roberts D. W., Staples R. C. Molecular cloning and regulatory analysis of the cuticle-degrading-protease structural gene from the entomopathogenic fungus Metarhizium anisopliae. Eur J Biochem. 1992 Mar 15;204(3):991–1001. doi: 10.1111/j.1432-1033.1992.tb16721.x. [DOI] [PubMed] [Google Scholar]
  26. Tang C. M., Cohen J., Krausz T., Van Noorden S., Holden D. W. The alkaline protease of Aspergillus fumigatus is not a virulence determinant in two murine models of invasive pulmonary aspergillosis. Infect Immun. 1993 May;61(5):1650–1656. doi: 10.1128/iai.61.5.1650-1656.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tatsumi H., Ogawa Y., Murakami S., Ishida Y., Murakami K., Masaki A., Kawabe H., Arimura H., Nakano E., Motai H. A full length cDNA clone for the alkaline protease from Aspergillus oryzae: structural analysis and expression in Saccharomyces cerevisiae. Mol Gen Genet. 1989 Oct;219(1-2):33–38. doi: 10.1007/BF00261154. [DOI] [PubMed] [Google Scholar]
  28. Toneguzzo F., Glynn S., Levi E., Mjolsness S., Hayday A. Use of a chemically modified T7 DNA polymerase for manual and automated sequencing of supercoiled DNA. Biotechniques. 1988 May;6(5):460–469. [PubMed] [Google Scholar]
  29. Tunlid A., Rosén S., Ek B., Rask L. Purification and characterization of an extracellular serine protease from the nematode-trapping fungus Arthrobotrys oligospora. Microbiology. 1994 Jul;140(Pt 7):1687–1695. doi: 10.1099/13500872-140-7-1687. [DOI] [PubMed] [Google Scholar]
  30. Watson M. E. Compilation of published signal sequences. Nucleic Acids Res. 1984 Jul 11;12(13):5145–5164. doi: 10.1093/nar/12.13.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES