Identification and characterization of a nuclear receptor subfamily I member in the Platyhelminth Schistosoma mansoni (SmNR1)
Identification and characterization of a nuclear receptor subfamily I member in the Platyhelminth Schistosoma mansoni (SmNR1)
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DOI：
10.1111/j.1742-4658.2006.05587.x
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发表时间：
2007-01-01
期刊：
影响因子：
5.4
通讯作者：
LoVerde, Philip T.
中科院分区：
文献类型：
--
作者：
Wu, Wenjie;Niles, Edward G.;LoVerde, Philip T.
一个cDNA编码的核受体亚家族I成员在扁形动物曼氏血吸虫（SmNR 1）的鉴定和表征。SmNR 1 cDNA全长2406 bp，含有一个开放阅读框，编码715个氨基酸残基的蛋白质。系统发育分析表明，SmNR 1是核受体亚家族I的一个分歧成员，没有已知的直系同源物。SmNR 1定位于S.荧光原位杂交检测Mansoni 1号染色体。SmNR 1的基因结构被确定，显示它由跨越超过14 kb的8个外显子组成。实时荧光定量RT-PCR结果表明，SmNR 1在整个虫体发育过程中均有表达，在卵、孢子囊和21日蠕虫中表达量较高。SmNR 1在A/B结构域中包含自主反式激活功能（AF 1），如酵母单杂交试验所示;它在酵母双杂交试验和谷胱甘肽S-转移酶下拉试验中与SmRXR 1相互作用。电泳迁移率变动分析表明，SmNR 1与SmRXR 1形成异源二聚体，与含有AGGTCA半位点、相隔0-5个核苷酸的半位点直接重复序列（DR 1 ~ DR 5）和未被核酸分隔的半位点回文重复序列（Pal 0）的DNA元件结合。瞬时转染哺乳动物COS-7细胞表明，SmNR 1/SmRXR 1可以增强DR 2依赖的报告基因的转录激活。我们的研究结果表明，SmNR 1是SmRXR 1的伙伴。
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.