Identification of a cDNA encoding a retinoid X receptor homologue from Schistosoma mansoni -: Evidence for a role in female-specific gene expression
Identification of a cDNA encoding a retinoid X receptor homologue from Schistosoma mansoni -: Evidence for a role in female-specific gene expression
复制标题
DOI：
10.1074/jbc.274.8.4577
复制
发表时间：
1999-02-19
影响因子：
4.8
通讯作者：
LoVerde, PT
中科院分区：
文献类型：
--
作者：
Freebern, WJ;Osman, A;LoVerde, PT
曼氏血吸虫是一种多细胞真核血吸虫，是世界范围内人类发病的主要原因。雌性寄生虫的生长，发育和基因调控的研究是很重要的，因为产生的卵是负责观察血吸虫病的发病机制，p14，蛋壳前体基因只在性成熟的女性在响应男性刺激，是女性特异性基因调控的模型。p14基因的上游区域与昆虫基因中已知的由类固醇受体超家族成员以性别、时间和组织特异性方式调节的序列相同。在此，我们报道了编码曼氏血吸虫（Sm）RXR同源物的cDNA的鉴定和表征。序列分析预测和Western印迹分析证实了74-kDa蛋白的合成，迄今为止报道的RXR家族中最大的成员。我们通过电泳迁移率变动分析表明，SmRXR结合p14基因的顺式元件，包括一个直接重复序列，遵循类固醇受体超家族成员识别的结合元件的“3-4-5”规则，此外，我们证明SmRXR可以作为酵母单杂交系统中的转录激活因子。通过定量逆转录-聚合酶链反应，我们表明，SmRXR基因的组成型表达，因此必须发挥多种作用，在整个生命周期的染色体。
Schistosoma mansoni, a multicelluar eukaryotic blood fluke, is a major cause of morbidity worldwide in humans. The study of female parasite growth, development, and gene regulation is important because the eggs produced are responsible for the pathogenesis observed in schistosomiasis, p14, an eggshell precursor gene expressed only in sexually mature females in response to a male stimulus, is a model for female-specific gene regulation. The upstream region of the p14 gene shares sequences present in insect genes known to be regulated in a sex-, temporal-, and tissue-specific manner by members of the steroid receptor superfamily, Herein, we report the identification and characterization of a cDNA that encodes the S, mansoni (Sm) RXR homologue, Sequence analysis predicts and Western blot analysis confirms the synthesis of a 74-kDa protein, the largest member of the RXR family reported to date. We show by electrophoretic mobility shift assay analysis that SmRXR binds to cis-elements of the p14 gene including a direct repeat that follows the "3-4-5" rule of binding elements recognized by members of the steroid receptor superfamily, Furthermore, we demonstrate that SmRXR can act as a transcription activator in the yeast one-hybrid system. Through quantitative reverse transcriptase-polymerase chain reaction, we show that the SmRXR gene is constitutively expressed and thus must play multiple roles throughout the schistosome life cycle.