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Regulation of cuticle-degrading subtilisin proteases from the entomopathogenic fungi, Lecanicillium spp: implications for host specificity

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Abstract

The ability to produce cuticle-degrading proteases to facilitate host penetration does not distinguish per se entomopathogenic fungi from saprophytes. However, adapted pathogens may produce host-protein specific enzymes in response to cues. This possibility prompted an investigation of the regulation of isoforms of the subtilisin Pr1-like proteases from five aphid-pathogenic isolates of Lecanicillium spp. Significant differences were found in substrate specificity and regulation of Pr1-like proteases between isoforms of the same isolate and between different isolates. For example, the pI 8.6 isoform from KV71 was considerably more active against aphid than locust cuticle and was induced specifically by N-acetylglucosamine (NAG). Isoform pI 9.1 from the same isolate was only produced on insect cuticle while most other isoforms were more prominent on chitin containing substrates but not induced by NAG. The ability to regulate isoforms independently may allow production at critical points in host penetration. Appearance of proteases (not subtilisins) with pI 4.2 and 4.4 only on aphid cuticle was a possible link with host specificity of KV71. The absence of C or N metabolite repression in subtilisins from KV42 is unusual for pathogen proteases and may help to account for differences in virulence strategy between aphid-pathogenic isolates of Lecanicillium longisporum (unpublished data).

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Abbreviations

dH2O:

Distilled water

Suc–Ala–Ala–Pro–Phe–NA:

Succinyl–alanyl–alanyl–prolyl–phenylalanyl–p–nitroanilide

Bz–Phe–Val–Arg–NA:

N–benzoyl–phenylalanyl–valyl–arginyl–p–nitroanilide

NAG:

N-acetylglucosamine

BSA:

Bovine serum albumen

IEF:

Isoelectric focusing

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Acknowledgements

This study was supported by CEC grant AIR3 CT94-1352 entitled “Insect pathogenic fungi for environmentally friendly pest control in the glasshouse”. We would to thank Chris Vennard for rearing the locusts and two anonymous reviewers whose comments on the initial submission significantly improved finished product.

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  1. Natasha J. Bye

    Present address: Nutricia Ltd, Trowbridge, BA14 0XQ, UK

Authors and Affiliations

  1. Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, BA2 7AY, UK

    Natasha J. Bye & A. Keith Charnley

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  1. Natasha J. Bye
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  2. A. Keith Charnley
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Correspondence to A. Keith Charnley.

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Communicated by Axel Brakhage.

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Bye, N.J., Charnley, A.K. Regulation of cuticle-degrading subtilisin proteases from the entomopathogenic fungi, Lecanicillium spp: implications for host specificity. Arch Microbiol 189, 81–92 (2008). https://doi.org/10.1007/s00203-007-0296-8

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  • DOI: https://doi.org/10.1007/s00203-007-0296-8

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