title: Molecular and functional characterisation of the heat shock protein 10 of Strongyloides ratti.
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.
authors: Bernhard Nocht Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany.
authors: Yasmina
authors: Y
authors: Tazir
authors: Vera
authors: V
authors: Steisslinger
authors: Hanns
authors: H
authors: Soblik
authors: Abuelhassan Elshazly
authors: AE
authors: Younis
authors: Svenja
authors: S
authors: Beckmann
authors: Christoph G
authors: CG
authors: Grevelding
authors: Hanno
authors: H
authors: Steen
authors: Norbert W
authors: NW
authors: Brattig
authors: Klaus D
authors: KD
authors: Erttmann
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D018835:Chaperonin 10 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D018834:Chaperonin 60 / Q000378:metabolism; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D019281:Dimerization; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D025941:Protein Interaction Mapping; D051381:Rats; D017208:Rats, Wistar; D017422:Sequence Analysis, DNA; D017170:Strongyloides ratti / Q000502:physiology*; D020798:Two-Hybrid System Techniques
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018835:Chaperonin 10; D018834:Chaperonin 60; D018509:DNA, Helminth