Wegrecki Marcin, Neira Jose Luis, Bravo Jeronimo
Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Avda. del Ferrocarril s/n, 03202 Elche (Alicante), Spain; Instituto de Biocomputación y Física de los Sistemas Complejos (BIFI), 50009 Zaragoza, Spain.
Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas, c/ Jaime Roig 11, 46010 Valencia, Spain.
PLoS One. 2015 Apr 16;10(4):e0123463. doi: 10.1371/journal.pone.0123463. eCollection 2015.
Erb1 (Eukaryotic Ribosome Biogenesis 1) protein is essential for the maturation of the ribosomal 60S subunit. Functional studies in yeast and mammalian cells showed that altogether with Nop7 and Ytm1 it forms a stable subcomplex called PeBoW that is crucial for a correct rRNA processing. The exact function of the protein within the process remains unknown. The N-terminal region of the protein includes a well conserved region shown to be involved in PeBoW complex formation whereas the carboxy-terminal half was predicted to contain seven WD40 repeats. This first structural report on Erb1 from yeast describes the architecture of a seven-bladed β-propeller domain that revealed a characteristic extra motif formed by two α-helices and a β-strand that insert within the second WD repeat. We performed analysis of molecular surface and crystal packing, together with multiple sequence alignment and comparison of the structure with other β-propellers, in order to identify areas that are more likely to mediate protein-protein interactions. The abundance of many positively charged residues on the surface of the domain led us to investigate whether the propeller of Erb1 might be involved in RNA binding. Three independent assays confirmed that the protein interacted in vitro with polyuridilic acid (polyU), thus suggesting a possible role of the domain in rRNA rearrangement during ribosome biogenesis.
Erb1(真核生物核糖体生物合成1)蛋白对于核糖体60S亚基的成熟至关重要。在酵母和哺乳动物细胞中的功能研究表明,它与Nop7和Ytm1共同形成一个名为PeBoW的稳定亚复合物,这对于正确的rRNA加工至关重要。该蛋白在此过程中的具体功能尚不清楚。该蛋白的N端区域包括一个保守性良好的区域,已证明该区域参与PeBoW复合物的形成,而其羧基末端的一半预计包含七个WD40重复序列。这份来自酵母的关于Erb1的首个结构报告描述了一个七叶β-螺旋桨结构域的结构,该结构域揭示了一个由两个α-螺旋和一条β-链形成的独特额外基序,该基序插入第二个WD重复序列中。我们进行了分子表面和晶体堆积分析,以及多序列比对,并将该结构与其他β-螺旋桨进行比较,以确定更有可能介导蛋白质-蛋白质相互作用的区域。该结构域表面大量带正电荷的残基促使我们研究Erb1的螺旋桨是否可能参与RNA结合。三项独立实验证实该蛋白在体外与聚尿苷酸(polyU)相互作用,从而表明该结构域在核糖体生物合成过程中rRNA重排中可能发挥的作用。