Isolation and characterization of Schistosoma mansoni constitutive androstane receptor

Rong Hu, Wenjie Wu, Edward G. Niles, Philip T. LoVerde

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

Full length cDNA clones encoding a Schistosoma mansoni homologue of vertebrate CAR/PXR/VDR group nuclear receptor, termed SmCAR were isolated from screening a S. mansoni adult worm cDNA library. SmCAR is a 702 amino acid protein which retains a typical domain organization of nuclear receptor superfamily members. A homology search demonstrated that SmCAR exhibits the highest homology with mouse constitutive androstane receptor (CAR). Like its orthologues from invertebrates, SmCAR contains a P box sequence of ESCKA in the DNA binding domain. The P box is important in determining the DNA binding specificity for nuclear receptors. SmCAR mRNA is expressed in every stage of S. mansoni life cycle with an elevated expression level in egg and cercaria stages. Two forms (78 and 81 kDa) of SmCAR protein were detected in schistosome worm extract by Western blot analysis. SmCAR protein was demonstrated to be widely distributed in adult worms by immunolocalization studies, being found in the subtegument in both male and female worms and in the ovaries, vitellaria and eggs in female worms. In vitro DNA binding assays demonstrated that SmCAR binds to the hsp27 ecdysone response element (EcRE) as well as schistosome p14 gene upstream region. The AGTGCA half site is essential for binding of SmCAR to the p14 gene upstream region. Therefore, AGTGCA probably serves as a high affinity binding half site for ESCKA containing nuclear receptors.

Original languageEnglish (US)
Pages (from-to)31-43
Number of pages13
JournalMolecular and Biochemical Parasitology
Volume148
Issue number1
DOIs
StatePublished - Jul 1 2006

Keywords

  • Constitutive androstane receptor (CAR)
  • Ecdysone response element (EcRE)
  • Nuclear receptor
  • Schistosoma mansoni
  • p14 female-specific gene

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

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