Isolation and characterization of Schistosoma mansoni constitutive androstane receptor.
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.
Hu
Rong
R
Department of Microbiology and Immunology, School of Medicine, State University of New York, Buffalo, NY 14214, USA.
Wu
Wenjie
W
Niles
Edward G
EG
Loverde
Philip T
PT
D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D000090702:Constitutive Androstane Receptor; D018509:DNA, Helminth / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005260:Female; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010053:Ovary / Q000378:metabolism; D010063:Ovum / Q000378:metabolism; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012550:Schistosoma mansoni / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D000090702:Constitutive Androstane Receptor; D018509:DNA, Helminth; D004268:DNA-Binding Proteins; D015801:Helminth Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors