SELECTION OF A SUBSET OF MESSENGER-RNAS FROM COMBINATORIAL 3' UNTRANSLATED REGION LIBRARIES USING NEURONAL RNA-BINDING PROTEIN HEL-N1
SELECTION OF A SUBSET OF MESSENGER-RNAS FROM COMBINATORIAL 3' UNTRANSLATED REGION LIBRARIES USING NEURONAL RNA-BINDING PROTEIN HEL-N1
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DOI：
10.1073/pnas.91.23.11207
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发表时间：
1994-11-08
影响因子：
11.1
通讯作者：
KEENE, JD
中科院分区：
文献类型：
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作者：
GAO, FB;CARSON, CC;KEENE, JD
Hel-N1是一种人类RNA结合蛋白，与果蝇神经元发育所必需的蛋白ELAV具有显著的同源性。Hel-N1已被证明在体外结合到编码c-myc、c-fos、粒细胞/巨噬细胞集落刺激因子和转录抑制因子的mRNA的3'非翻译区（同上）。我们报告Hel-N1和相关形式Hel-N2在人髓母细胞瘤细胞中表达，但它们的比例与成人脑和胎儿脑中的比例显著不同。从随机化组合文库中选择RNA靶标产生了Hel-N1和Hel-N2的（A+U）丰富的共有序列。作为鉴定这些蛋白质的细胞RNA靶标的手段，我们设计了代表天然来源的3'非翻译区的组合形状文库，并且能够选择与Hel-N1结合的转录物的结构相关子集。大约10%的蛋白质编码的这些子集mRNA的数据库中是可识别的，大多数涉及细胞生长调节。这种方法提供了一种通过使用RNA结合蛋白分配含有共同序列元件的mRNA来获得在各种细胞类型中表达的新基因的手段。
Hel-N1, a human RNA binding protein, shares significant homology with Drosophila protein ELAV, which is essential for fly neuronal development. Hel-N1 has been shown to bind in vitro to 3' untranslated regions of mRNAs encoding c-myc, c-fos, granulocyte/macrophage colony-stimulating factor, and transcriptional repressor, Id. We report that Hel-N1 and a related form, Hel-N2,are expressed in human medulloblastoma cells, but their ratio differs significantly from that in adult brain and fetal brain. Selection of RNA targets from randomized combinatorial libraries yielded (A+U)-rich consensus sequences for both Hel-N1 and Hel-N2. As a means to identify cellular RNA targets for these proteins, we devised combinatorial shape libraries representing naturally derived 3' untranslated regions and were able to select a structurally related subset of transcripts that bound to Hel-N1. Approximately 10% of the proteins encoded by these subset mRNAs were identifiable in the data bases and most are implicated in cell growth regulation. This approach provides a means to gain access to novel genes expressed in various cell types by partitioning mRNAs containing common sequence elements using RNA-binding proteins.