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Identification and characterization of a nuclear receptor subfamily I member in the Platyhelminth Schistosoma mansoni (SmNR1)
Author(s) -
Wu Wenjie,
Niles Edward G,
Hirai Hirohisa,
LoVerde Philip T
Publication year - 2007
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2006.05587.x
Subject(s) - biology , subfamily , microbiology and biotechnology , complementary dna , schistosoma mansoni , cdna library , transactivation , gene , reporter gene , genetics , gene expression , zoology , helminths , schistosomiasis
A cDNA encoding a nuclear receptor subfamily I member in the platyhelminth Schistosoma mansoni (SmNR1) was identified and characterized. SmNR1 cDNA is 2406 bp long and contains an open reading frame encoding a 715 residue protein. Phylogenetic analysis demonstrates that SmNR1 is a divergent member of nuclear receptor subfamily I with no known orthologue. SmNR1 was localized to S. mansoni chromosome 1 by fluorescent in situ hybridization. Gene structure of SmNR1 was determined showing it to consist of eight exons spanning more than 14 kb. Quantitative real‐time RT‐PCR showed that SmNR1 was expressed throughout schistosome development with a higher expression in eggs, sporocysts and 21‐day worms. SmNR1 contains an autonomous transactivation function (AF1) in the A/B domain as demonstrated in a yeast one‐hybrid assay; it interacts with SmRXR1 in a yeast two‐hybrid assay and in a glutathione S ‐transferase pull‐down assay. Electrophoretic mobility shift assay showed that SmNR1 could form a heterodimer with SmRXR1 to bind to DNA elements containing the half‐site AGGTCA, a direct repeat of the half‐site separated by 0–5 nucleotides (DR1‐DR5) and a palindrome repeat of the half‐site not separated by nucleic acids (Pal0). Transient transfection in mammalian COS‐7 cells showed that SmNR1/SmRXR1 could enhance the transcriptional activation of a DR2‐dependent reporter gene. Our results demonstrate that SmNR1 is a partner of SmRXR1.