Molecular and functional characterisation of the heat shock protein 10 of Strongyloides ratti

Mol Biochem Parasitol. 2009 Dec;168(2):149-57. doi: 10.1016/j.molbiopara.2009.07.007. Epub 2009 Jul 28.

Abstract

Strongyloides stercoralis and S. ratti are intestinal parasitic nematodes infecting rats and humans, respectively. Both present extraordinary life cycles comprising a free-living generation in addition to parasitic stages. In search of molecules possibly involved in parasite-host interaction, we performed mass spectrometry to identify excretory/secretory products of S. ratti. Amongst others we detected homologs of the heat shock proteins HSP10 and HSP60 (Sr-HSP10 and Sr-HSP60). HSPs are well known as chaperones involved in stress responses of cells, but recent studies suggest additional roles of small HSPs for parasite biology including immune modulation. To characterise Sr-HSP10, we cloned its full-length cDNA, analysed the genomic organisation, tested its presumptive role as an interaction partner of Sr-HSP60, studied its transcription in the parasite, and expressed the protein to test its immune responses. The cDNA contains an open reading frame of 330bp encoding a polypeptide of 110 amino acids with an approximate molecular weight of 10kDa. The Sr-HSP10 protein is highly homologous to that of the human pathogen S. stercoralis with only eight amino acid substitutions. Analysis of the genomic organisation of the Sr-HSP10 locus revealed that the gene is linked head-to-head to the gene encoding Sr-HSP60, and both share a bidirectional promoter. RT-PCR experiments indicated potential independent expression of the Sr-HSPs genes. In situ hybridisation results demonstrate Sr-HSP10 transcription in the gut area. Mammalian and yeast two-hybrid assays show dimerisation of Sr-HSP10, but no binding to recombinant Sr-HSP60. Immunisation experiments finally revealed a strong immunogenicity of Sr-HSP10 and provided evidence for a role in regulating the host-parasite interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Helminth / blood
  • Antigens, Helminth / chemistry
  • Antigens, Helminth / genetics
  • Antigens, Helminth / immunology
  • Antigens, Helminth / metabolism
  • Base Sequence
  • Chaperonin 10 / chemistry
  • Chaperonin 10 / genetics*
  • Chaperonin 10 / immunology
  • Chaperonin 10 / metabolism*
  • Chaperonin 60 / metabolism
  • DNA, Helminth / chemistry
  • DNA, Helminth / genetics
  • Dimerization
  • Gene Expression Profiling
  • Molecular Sequence Data
  • Molecular Weight
  • Protein Binding
  • Protein Interaction Mapping
  • Rats
  • Rats, Wistar
  • Sequence Analysis, DNA
  • Strongyloides ratti / physiology*
  • Two-Hybrid System Techniques

Substances

  • Antibodies, Helminth
  • Antigens, Helminth
  • Chaperonin 10
  • Chaperonin 60
  • DNA, Helminth

Associated data

  • GENBANK/FJ694974