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The Tripartite Motif of Nuclear Factor 7 Is Required for Its Association with Transcriptional Units

机译：需要核因子7的三方基序与转录单位结合。

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In amphibian oocytes, the maternal nuclear factor NF7 associates with the elongating pre-mRNAs present on the numerous lateral loops of the lampbrush chromosomes. Here, we have purified NF7 from an oocyte extract by using a combination of ion-exchange chromatography and gel filtration chromatography and demonstrated for the first time that nucleoplasmic NF7 exists primarily as free homotrimers. We confirmed the in vivo homotrimerization of NF7 by using a glutaraldehyde cross-linking assay, and we further showed that it only requires the coiled-coil domain of the NF7 tripartite motif/RBCC motif. Interestingly, we also obtained evidence that NF7 is recruited to the nucleus as a homotrimer, and expression of several mutated forms of NF7 in oocytes demonstrated that both the coiled coil and B box of NF7 are required for its chromosomal association. Together, these data strongly suggest that the interaction of NF7 with the active transcriptional units of RNA polymerase II is mediated by a trimeric B box. Finally, and in agreement with a role for NF7 in pre-mRNA maturation, we obtained evidence supporting the idea that NF7 associates with Cajal bodies.
机译：在两栖卵母细胞中，母体核因子NF7与存在于笔刷染色体众多侧向环中的延长前mRNA缔合。在这里，我们通过结合使用离子交换色谱和凝胶过滤色谱从卵母细胞提取物中纯化NF7，并首次证明核质NF7主要以游离同三聚体的形式存在。我们通过使用戊二醛交联测定法证实了NF7的体内均三聚化，并且我们进一步表明，它仅需要NF7三方基序/ RBCC基序的卷曲螺旋结构域。有趣的是，我们还获得了证据表明NF7以同源三聚体的形式被募集到细胞核中，并且在卵母细胞中表达了几种突变形式的NF7，这证明了线圈的卷曲和NF7的B盒都需要其染色体结合。总之，这些数据强烈暗示NF7与RNA聚合酶II的活性转录单位的相互作用是由三聚体B框介导的。最后，与NF7在mRNA之前成熟中的作用相一致，我们获得了支持NF7与Cajal体相关联的观点的证据。

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