Poly (ADP-ribose) polymerase-1 (PARP-1) is a multifunctional protein that is associated with various biological processes like chromatin remodeling, DNA damage, cell death etc. In Dictyostelium discoideum, PARP-1 has also been implicated in cellular differentiation and development. However, its interacting proteins during multicellular development are not yet explored. Hence, the present study aims to identify PARP-1 interacting proteins during multicellular development of D. discoideum.

BRCA1 C-terminus (BRCT) domain of PARP-1, which is mainly involved in protein-protein interactions was cloned in pGEX4T1 vector and developmental interactome of PARP-1 were analyzed by affinity purification-mass spectrometry. These interactions were further confirmed by in-silico protein-protein docking analysis, which led to identification of the proteins that show high affinity for BRCT domain. Initially, the protein structures were modeled on SWISS MODEL and PHYRE2 servers, refined by 3Drefine and validated by PROCHECK. Further, interaction sites of BRCT and the conserved regions in all interacting proteins were predicted using cons-PPISP and ConSurf, respectively. Finally, protein-protein docking analysis was done by HADDOCK.

Our results identified 19 possible BRCT interacting proteins during D. discoideum development. Furthermore, interacting residues involved in the interactions and functional regions were explored. This is the first report where PARP-1's developmental interactome in D. discoideum is well established. The current findings demonstrate PARP-1's developmental interactome in D. discoideum and provide the groundwork to understand its regulated functions in developmental biology which would undoubtedly extend our perception towards developmental diseases in higher complex organisms and their treatment.

聚 (ADP-核糖) 聚合酶-1 (PARP-1) 是一种多功能蛋白质，与染色质重塑、DNA 损伤、细胞死亡等各种生物过程相关。在盘基网柄菌中，PARP-1 也与细胞分化有关和发展。然而，尚未探索其在多细胞发育过程中的相互作用蛋白。因此，本研究旨在鉴定D. discoideum 多细胞发育过程中的 PARP-1 相互作用蛋白。

将主要参与蛋白质-蛋白质相互作用的PARP-1的BRCA1 C-末端（BRCT）结构域克隆到pGEX4T1载体中，并通过亲和纯化-质谱分析PARP-1的发育相互作用组。通过计算机内蛋白质-蛋白质对接分析进一步证实了这些相互作用，从而鉴定出对 BRCT 结构域具有高亲和力的蛋白质。最初，蛋白质结构在 SWISS MODEL 和 PHYRE2 服务器上建模，由 3Drefine 改进并由 PROCHECK 验证。此外，分别使用 cons-PPISP 和 ConSurf 预测了 BRCT 的相互作用位点和所有相互作用蛋白中的保守区域。最后，蛋白质-蛋白质对接分析由HADDOCK完成。

我们的结果在D. discoideum发育过程中确定了 19 种可能的 BRCT 相互作用蛋白。此外，探索了参与相互作用和功能区域的相互作用残基。这是第一份报告，其中 PARP-1 在D. discoideum中的发育相互作用组得到了很好的证实。目前的研究结果证明了 PARP-1 在D. discoideum中的发育相互作用组，并为了解其在发育生物学中的调节功能奠定了基础，这无疑将扩展我们对高级复杂生物体发育疾病及其治疗的认识。