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首页> 外文期刊>Nucleic Acids Research >RNA recognition and self-association of CPEB4 is mediated by its tandem RRM domains
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RNA recognition and self-association of CPEB4 is mediated by its tandem RRM domains

机译：CPEB4的RNA识别和自我关联由其串联RRM域介导

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摘要

Cytoplasmic polyadenylation is regulated by the interaction of the cytoplasmic polyadenylation element binding proteins (CPEB) with cytoplasmic polyadenylation element (CPE) containing mRNAs. The CPEB family comprises four paralogs, CPEB1-4, each composed of a variable N-terminal region, two RNA recognition motif (RRM) and a C-terminal ZZ-domain. We have characterized the RRM domains of CPEB4 and their binding properties using a combination of biochemical, biophysical and NMR techniques. Isothermal titration calorimetry, NMR and electrophoretic mobility shift assay experiments demonstrate that both the RRM domains are required for an optimal CPE interaction and the presence of either one or two adenosines in the two most commonly used consensus CPE motifs has little effect on the affinity of the interaction. Both the single RRM1 and the tandem RRM1-RRM2 have the ability to dimerize, although representing a minor population. Self-association does not affect the proteins' ability to interact with RNA as demonstrated by ion mobility-mass spectrometry. Chemical shift effects measured by NMR of the apo forms of the RRM1-RRM2 samples indicate that the two domains are orientated toward each other. NMR titration experiments show that residues on the beta-sheet surface on RRM1 and at the C-terminus of RRM2 are affected upon RNA binding. We propose a model of the CPEB4 RRM1-RRM2-CPE complex that illustrates the experimental data.
机译：细胞质多腺苷酸化通过细胞质多腺苷酸聚合物结合蛋白（CPEB）与含MRNA的细胞质多腺苷酸化元素（CPE）的相互作用来调节。 CPEB系列包括四个副病虫戈，CPEB1-4，各自由可变N末端区域，两个RNA识别基序（RRM）和C末端ZZ域组成。我们使用生物化学，生物物理和NMR技术的组合表征了CPEB4的RRM结构域及其结合特性。等温滴定热量测定法，NMR和电泳迁移率偏移测定实验表明，最佳CPE相互作用所需的是，两种最常用的共识CPE基序中的一个或两个腺苷的存在对其的亲和力几乎没有影响相互作用。单个RRM1和串联RRM1-RRM2都具有二聚体的能力，尽管代表轻微的人口。自我关联不会影响蛋白质与RNA相互作用的能力，如离子迁移率质谱法所示。通过RRM1-RRM2样品的APO形式测量的化学换档效果表明两个域以彼此为定向。 NMR滴定实验表明RRM1上β-片材表面上的残留物和RRM2的C末端的残留物受RNA结合的影响。我们提出了一种模型，用于说明实验数据的CPEB4 RRM1-RRM2-CPE复合体。

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