Department of Biological and Geographical Sciences, School of Applied Sciences, University of Huddersfield, Huddersfield, UK.
Protein Sci. 2021 Jun;30(6):1196-1209. doi: 10.1002/pro.4085. Epub 2021 May 10.
Polymerase δ-interacting protein 2 (POLDIP2, PDIP38) is a multifaceted, "moonlighting" protein, involved in binding protein partners from many different cellular processes, including mitochondrial metabolism and DNA replication and repair. How POLDIP2 interacts with many different proteins is unknown. Towards this goal, we present the crystal structure of POLDIP2 to 2.8 Å, which exhibited a compact two-domain β-strand-rich globular structure, confirmed by circular dichroism and small angle X-ray scattering approaches. POLDIP2 comprised canonical DUF525 and YccV domains, but with a conserved domain linker packed tightly, resulting in an "extended" YccV module. A central channel was observed, which we hypothesize could influence structural changes potentially mediated by redox conditions, following observation of a modified cysteine residue in the channel. Unstructured regions were rebuilt by ab initio modelling to generate a model of full-length POLDIP2. Molecular dynamics simulations revealed a highly dynamic N-terminal region tethered to the YccV-domain by an extended linker, potentially facilitating interactions with distal binding partners. Models of POLDIP2 complexed with two of its partners, PrimPol and PCNA, indicated that dynamic flexibility of the POLDIP2 N-terminus and loop regions likely mediate protein interactions.
聚合酶 δ 相互作用蛋白 2(POLDIP2,PDIP38)是一种多方面的“兼职”蛋白,参与结合来自许多不同细胞过程的蛋白伴侣,包括线粒体代谢、DNA 复制和修复。目前尚不清楚 POLDIP2 如何与许多不同的蛋白质相互作用。为此,我们呈现了 POLDIP2 的 2.8Å 晶体结构,该结构表现出紧凑的双域 β-折叠丰富的球状结构,通过圆二色性和小角 X 射线散射方法得到了证实。POLDIP2 包含典型的 DUF525 和 YccV 结构域,但保守的结构域接头紧密包装,导致“扩展”的 YccV 模块。观察到通道中的修饰半胱氨酸残基后,我们假设可以观察到中央通道,这可能会影响潜在由氧化还原条件介导的结构变化。无规则区域通过从头建模进行重建,以生成全长 POLDIP2 的模型。分子动力学模拟揭示了一个高度动态的 N 端区域通过延伸的接头与 YccV 结构域连接,这可能有利于与远端结合伴侣的相互作用。与两个 POLDIP2 伴侣 PrimPol 和 PCNA 形成复合物的模型表明,POLDIP2 N 端和环区的动态灵活性可能介导蛋白相互作用。