The gene for histone RNA hairpin binding protein is located on human chromosome 4 and encodes a novel type of RNA binding protein
The gene for histone RNA hairpin binding protein is located on human chromosome 4 and encodes a novel type of RNA binding protein
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DOI：
10.1093/emboj/16.4.769
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发表时间：
1997-02-17
期刊：
影响因子：
11.4
通讯作者：
Muller, B
中科院分区：
文献类型：
--
作者：
Martin, F;Schaller, A;Muller, B
动物组蛋白mRNA 3'端的发夹结构控制组蛋白RNA的3'加工、核质转运、翻译和稳定性。在细胞核和多聚核糖体提取物中，已经鉴定出与组蛋白RNA发夹结合的蛋白质在功能上重叠（如果不相同的话）。我们自己的研究结果表明，这些发夹结合蛋白（HBPs）结合它们的目标RNA作为单体，所得的核糖核蛋白复合物是非常稳定的。这些特征促使我们在酿酒酵母中通过RNA介导的三杂交选择来选择HBP编码的人类cDNA。从一个选定的克隆的全细胞提取物含有Gal4融合蛋白，其与组蛋白发夹RNA以序列和结构特异性方式相互作用，类似于牛HBP富集的级分，表明cDNA编码HBP。DNA序列分析表明，编码序列不包含任何已知的RNA结合基序。HBP基因由8个外显子组成，位于4号染色体短臂上，全长19.5kb。HBP开放阅读框在体外的翻译产生了一个43 kDa的蛋白质，其RNA结合特异性与鼠或牛HBP相同。此外，在酿酒酵母中表达的重组HBP在组蛋白前体mRNA加工中具有功能，证实我们确实鉴定了人HBP基因。
The hairpin structure at the 3' end of animal histone mRNAs controls histone RNA 3' processing, nucleocytoplasmic transport, translation and stability of histone mRNA. Functionally overlapping, if not identical, proteins binding to the histone RNA hairpin have been identified in nuclear and polysomal extracts. Our own results indicated that these hairpin binding proteins (HBPs) bind their target RNA as monomers and that the resulting ribonucleoprotein complexes are extremely stable. These features prompted us to select for HBP-encoding human cDNAs by RNA-mediated three-hybrid selection in Saccharomyces cerevesiae. Whole cell extract from one selected clone contained a Gal4 fusion protein that interacted with histone hairpin RNA in a sequence- and structure-specific manner similar to a fraction enriched for bovine HBP, indicating that the cDNA encoded HBP. DNA sequence analysis revealed that the coding sequence did not contain any known RNA binding motifs. The HBP gene is composed of eight exons covering 19.5 kb on the short arm of chromosome 4. Translation of the HBP open reading frame in vitro produced a 43 kDa protein with RNA binding specificity identical to murine or bovine HBP. In addition, recombinant HBP expressed in S.cerevisiae was functional in histone pre-mRNA processing, confirming that we have indeed identified the human HBP gene.