Structural and Mechanistic Insights into Poly(uridine) Tract Recognition by the hnRNP C RNA Recognition Motif
Structural and Mechanistic Insights into Poly(uridine) Tract Recognition by the hnRNP C RNA Recognition Motif
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DOI：
10.1021/ja507690d
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发表时间：
2014-10-15
影响因子：
15
通讯作者：
Maris, Christophe
中科院分区：
文献类型：
--
作者：
Cienikova, Zuzana;Damberger, Fred F.;Maris, Christophe
HnRNP C是一种普遍存在的RNA调节因子，是核hnRNP核心颗粒的主要成分。该蛋白质含有一个氨基末端RNA识别基序（RRM），已知可结合富含尿苷（U）的序列。这项工作提供了这种相互作用的分子和机制的理解。我们解决了溶液结构的RRM在复杂的聚（U）寡聚体的五个和七个核苷酸。RRM的五个结合口袋以不寻常的5 '到3'碱基选择性梯度识别尿苷。因此，目标识别对碱基聚类非常敏感，这解释了对连续尿苷束的偏好。使用一种新的方法，将其多寄存器结合的热力学描述的RRM的结构衍生的识别共识，我们模拟了这种蛋白质的细胞尿苷道的饱和度。结合模式是显着一致的实验观察到的转录组范围内的全长hnRNP C短尿苷束的交联分布。这一结果重新确立了RRM作为hnRNP C四聚体的主要RNA结合结构域，并为使用结构方法解释高通量相互作用数据提供了概念证明。
HnRNP C is a ubiquitous RNA regulatory factor and the principal constituent of the nuclear hnRNP core particle. The protein contains one amino-terminal RNA recognition motif (RRM) known to bind uridine (U)-rich sequences. This work provides a molecular and mechanistic understanding of this interaction. We solved the solution structures of the RRM in complex with poly(U) oligomers of five and seven nucleotides. The five binding pockets of RRM recognize uridines with an unusual 5'-to-3' gradient of base selectivity. The target recognition is therefore strongly sensitive to base clustering, explaining the preference for contiguous uridine tracts. Using a novel approach integrating the structurally derived recognition consensus of the RRM with a thermodynamic description of its multi-register binding, we modeled the saturation of cellular uridine tracts by this protein. The binding pattern is remarkably consistent with the experimentally observed transcriptome-wide cross-link distribution of the full-length hnRNP C on short uridine tracts. This result re-establishes the RRM as the primary RNA-binding domain of the hnRNP C tetramer and provides a proof of concept for interpreting high-throughput interaction data using structural approaches.