Shah Binita, Hunkeler Moritz, Bratt Ariana, Yue Hong, Jaen Maisonet Isabella, Fischer Eric S, Buhrlage Sara J
Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA, USA.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
Nat Commun. 2025 Aug 28;16(1):8025. doi: 10.1038/s41467-025-63161-3.
VCP/p97 regulates a wide range of cellular processes, including post-mitotic Golgi reassembly. In this context, VCP is assisted by p47, an adapter protein, and VCPIP1, a deubiquitylase (DUB). However, how they organize into a functional ternary complex to promote Golgi assembly remains unknown. Here, we use cryo-EM to characterize both VCP-VCPIP1 and VCP-VCPIP1-p47 complexes. We show that VCPIP1 engages VCP through two interfaces: one involving the N-domain of VCP and the UBX domain of VCPIP1, and the other involving the VCP D2 domains and a region of VCPIP1 we refer to as VCPID. The p47 UBX domain competitively binds to the VCP N-domain, while not affecting VCPID binding. We show that VCPID is critical for VCP-mediated enhancement of DUB activity and proper Golgi assembly. The ternary structure along with biochemical and cellular data provides new insights into the complex interplay of VCP with its co-factors.
VCP/p97调控多种细胞过程，包括有丝分裂后高尔基体的重新组装。在这种情况下，VCP由衔接蛋白p47和去泛素化酶（DUB）VCPIP1协助。然而，它们如何组装成功能性三元复合物以促进高尔基体组装仍不清楚。在这里，我们使用冷冻电镜来表征VCP-VCPIP1和VCP-VCPIP1-p47复合物。我们发现VCPIP1通过两个界面与VCP结合：一个涉及VCP的N结构域和VCPIP1的UBX结构域，另一个涉及VCP的D2结构域和我们称为VCPID的VCPIP1区域。p47的UBX结构域竞争性结合VCP的N结构域，同时不影响VCPID的结合。我们表明VCPID对于VCP介导的DUB活性增强和高尔基体的正常组装至关重要。三元结构以及生化和细胞数据为VCP与其辅助因子之间复杂的相互作用提供了新的见解。
