The ubiquitin–proteasome system in Strongyloididae. Biochemical evidence for developmentally regulated proteolysis in Strongyloides venezuelensis

Abstract

Nematode parasites from the genus Strongyloides spp. are important pathogens of the intestinal mucosa of animals and humans. Their complex life cycles involve alternating developmental adaptations between larvae stages and the adult parthenogenetic female. Here, we report, primarily through homology-based searching, the existence of the major components of the ubiquitin–proteasome system in this genus, using the available EST data from S. ratti, S. stercoralis, and Parastrongyloides trichosuri. In this study, S. venezuelensis was used as our model organism for detection of proteasome activity and ubiquitinated substrates in cytosolic preparations from the L3 larvae and the adult female. Marked differences in proteasome capabilities were found when these two stages were compared. A preference for degradation of chymotryptic synthetic peptides was found in both stages with the adult exhibiting a higher rate of hydrolysis compared to the larvae. Due to the high evolutionary conservation of proteasome alpha subunits, an anti-human proteasome antibody was able to recognize proteasome subunits in these preparations by Western blotting, supporting the proposal that the activity of the ubiqutin–proteasome system is developmentally regulated in this nematode.

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Fig. 1

Abbreviations

DMSO:

Dimethyl sulfoxide

ESTs:

Expressed sequence tags

PBS:

Phosphate-buffered saline

DTT:

Dithiothreitol

1D:

One-dimensional gel electrophoresis

2D:

Two-dimensional gel electrophoresis

BCIP:

5-Bromo-4-chloro-3′-indolyphosphate p-toluidine salt

NBT:

Nitro-blue tetrazolium chloride

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Acknowledgements

This work was sponsored by the Conselho Nacional de Pesquisa e Desenvolvimento (CNPq). The authors thank Joao B. A de Oliveira (Unicamp—SP) for his expertise with parasite production and maintenance.

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Correspondence to Fabiana M. de Paula.

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de Paula, F.M., Castro-Borges, W., Júnior, O.S.P. et al. The ubiquitin–proteasome system in Strongyloididae. Biochemical evidence for developmentally regulated proteolysis in Strongyloides venezuelensis . Parasitol Res 105, 567 (2009). https://doi.org/10.1007/s00436-009-1430-0

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Keywords

  • Parasitic Nematode
  • Proteasome Activity
  • Proteasome Pathway
  • Antibody Binding Site
  • Parthenogenetic Female