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随机磷酸化的聚苯乙烯衍生物与RNA聚合酶II转录因子的相互作用:第一部分。

Randomly phosphorylated polystyrene derivatives interact with RNA polymerase II transcription factors: part I.

作者信息

Imbert E, Letourneur D, Jozefowicz M

机构信息

Laboratoire de Recherches sur les Macromolecules, CNRS URA 502, University Paris-Nord, Villetaneuse, France.

出版信息

J Biomed Mater Res. 1997 Mar 15;34(4):417-25. doi: 10.1002/(sici)1097-4636(19970315)34:4<417::aid-jbm3>3.0.co;2-n.

Abstract

Insoluble functional synthetic random copolymers are able to develop at their surfaces specific interactions with biologic components. Crosslinked phosphorylated polystyrene derivatives were previously shown to mimic DNA antigen because they interacted with anti-DNA antibodies found in the sera of systemic lupus erythematosus patients. These biospecific surfaces were postulated to be able to bind other DNA-binding proteins such as RNA polymerase II transcription factors. Indeed, these proteins play a major role in gene regulation in mammalian cells. This hypothesis was checked by adsorption and elution of HeLa cell nuclear extracts on a 72% phosphorylated resin. The composition of the eluted fractions were analyzed by electrophoresis, and the biologic activity of the transcription factors was tested using an in vitro transcription assay. The results showed that USF, TATA-binding protein (TBP), and TFIIB were specifically adsorbed on the polymer and that all eluted factors kept their biologic activity. Therefore, randomly phosphorylated polystyrene derivatives may be useful for the fractionation of RNA polymerase II transcription factors.

摘要

不溶性功能性合成无规共聚物能够在其表面与生物成分发生特异性相互作用。先前已表明,交联的磷酸化聚苯乙烯衍生物能够模拟DNA抗原,因为它们能与系统性红斑狼疮患者血清中发现的抗DNA抗体相互作用。据推测,这些生物特异性表面能够结合其他DNA结合蛋白,如RNA聚合酶II转录因子。实际上,这些蛋白质在哺乳动物细胞的基因调控中发挥着主要作用。通过在72%磷酸化树脂上吸附和洗脱HeLa细胞核提取物来验证这一假设。通过电泳分析洗脱组分的组成,并使用体外转录试验测试转录因子的生物活性。结果表明,上游刺激因子(USF)、TATA结合蛋白(TBP)和转录因子IIB(TFIIB)被特异性吸附在聚合物上,并且所有洗脱的因子都保持了它们的生物活性。因此,随机磷酸化的聚苯乙烯衍生物可能有助于RNA聚合酶II转录因子的分级分离。

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