对EDEN-BP特异性RNA结合的序列要求分析。
An analysis of the sequence requirements of EDEN-BP for specific RNA binding.
作者信息
Bonnet-Corven Sylvie, Audic Yann, Omilli Francis, Osborne H Beverley
机构信息
UMR 6061 CNRS-Génétique et Développement, IFR 97, Université de Rennes 1, Faculté de Médecine, 2 Avenue Léon Bernard, CS 34317, 35043 Rennes Cedex, France.
出版信息
Nucleic Acids Res. 2002 Nov 1;30(21):4667-74. doi: 10.1093/nar/gkf586.
Abstract
EDEN-BP (embryo deadenylation element-binding protein) binds specifically to the EDEN motif in the 3'-untranslated regions of maternal mRNAs and targets these mRNAs for deadenylation and translational repression in Xenopus laevis embryos. EDEN-BP contains three RNA recognition motifs (RRMs) and is related to the elav family of RNA-binding proteins. In the present study we show that the two N-terminal RRMs of EDEN-BP are necessary for the interaction with EDEN as well as a part of the linker region (between RRM2 and RRM3). Using a band shift assay we show that two different complexes are formed according to the size and, therefore, the functional nature of the EDEN motif. Finally, we show that EDEN-BP can form a dimer in a two-hybrid assay. Accordingly, we suggest that the functional configuration of EDEN-BP is a dimer.
摘要
EDEN-BP（胚胎去腺苷酸化元件结合蛋白）特异性结合非洲爪蟾胚胎中母体mRNA 3'非翻译区的EDEN基序，并将这些mRNA作为去腺苷酸化和翻译抑制的靶点。EDEN-BP包含三个RNA识别基序（RRMs），与RNA结合蛋白的elav家族相关。在本研究中，我们表明EDEN-BP的两个N端RRMs以及连接区的一部分（RRM2和RRM3之间）对于与EDEN的相互作用是必需的。使用凝胶迁移试验，我们表明根据EDEN基序的大小以及功能性质会形成两种不同的复合物。最后，我们表明在双杂交试验中EDEN-BP可以形成二聚体。因此，我们认为EDEN-BP的功能构型是二聚体。
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