Structural characterization of human RPA70N association with DNA damage response proteins.
Structural characterization of human RPA70N association with DNA damage response proteins.
复制标题
作者：
Wu Y;Fu W;Zang N;Zhou C
异三聚体复制蛋白A(RPA)是真核生物中普遍存在的单链DNA结合蛋白，几乎参与DNA代谢的方方面面，尤其是DNA损伤反应。RPA70亚基的N端OB结构域(RPA70N)是RPA的主要蛋白质相互作用元件，可与20多种蛋白质结合。先前对含有P53、DNA2和PrimPol片段的RPA70N的结晶学研究表明，RPA70N与模拟单链DNA的两亲性多肽结合。核磁共振化学位移研究也为RPA70N残基与靶序列的相互作用提供了有价值的信息。然而，目前还不清楚RPA70N是如何识别和区分如此多样化的一组靶蛋白的。在这里，我们介绍了RPA70N与八种DNA损伤反应蛋白的多肽形成的复合体的高分辨晶体结构。结构表明，除了ssDNA模拟相互作用模式外，RPA70N还采用多种方式结合其伴侣。我们的结果促进了对RPA70N介导的DNA损伤反应蛋白募集的机制的理解。
The heterotrimeric Replication protein A (RPA) is the ubiquitous eukaryotic single-stranded DNA (ssDNA) binding protein and participates in nearly all aspects of DNA metabolism, especially DNA damage response. The N-terminal OB domain of the RPA70 subunit (RPA70N) is a major protein-protein interaction element for RPA and binds to more than 20 partner proteins. Previous crystallography studies of RPA70N with p53, DNA2 and PrimPol fragments revealed that RPA70N binds to amphipathic peptides that mimic ssDNA. NMR chemical-shift studies also provided valuable information on the interaction of RPA70N residues with target sequences. However, it is still unclear how RPA70N recognizes and distinguishes such a diverse group of target proteins. Here, we present high-resolution crystal structures of RPA70N in complex with peptides from eight DNA damage response proteins. The structures show that, in addition to the ssDNA mimicry mode of interaction, RPA70N employs multiple ways to bind its partners. Our results advance the mechanistic understanding of RPA70N-mediated recruitment of DNA damage response proteins.