Department of Pharmacology, Faculty of Pharmaceutical Sciences, Doshisha Women's College of Liberal Arts, Kyotanabe, Japan.
Genes Cells. 2022 Apr;27(4):305-312. doi: 10.1111/gtc.12926. Epub 2022 Feb 21.
Mono(ADP-ribosyl)ation and poly(ADP-ribosyl)ation are posttranslational modifications evolutionarily conserved in prokaryotes and eukaryotes. They entail transfer of one or more ADP-ribose moieties from NAD to acceptor proteins with the simultaneous release of nicotinamide. The resultant ADP-ribosylated acceptor proteins regulate diverse cellular functions. For instance, ADP-ribosyltransferase 1 (ART1) catalyzes mono(ADP-ribosyl)ation of arginine residues in Trim72, a protein specifically expressed in muscle cells and involved in cell membrane repair, which is enhanced upon its ADP-ribosylation. By contrast, the contribution made by ADP-ribosylation to membrane repair in epithelial cells remains unclear. In this study, we investigated the involvement of ADP-ribosylation in cell membrane repair in HEK293T and HeLa cells. We found that upon induction of membrane damage using streptolysin-O, poly(ADP-ribose) polymerase 1 (PARP1) catalyzed poly(ADP-ribosyl)ation. In scratch assays, inhibition of PARP1 activity using the nonspecific PARP inhibitor PJ34 or shRNA targeting PARP1 delayed wound healing, suggesting that PARP1-catalyzed poly(ADP-ribosyl)ation plays a key role in membrane repair in epithelial cells.
单（ADP-核糖基）化和多（ADP-核糖基）化是原核生物和真核生物中进化保守的翻译后修饰。它们涉及将一个或多个 ADP-核糖基部分从 NAD 转移到具有同时释放烟酰胺的受体蛋白。由此产生的 ADP-核糖基化受体蛋白调节多种细胞功能。例如，ADP-核糖基转移酶 1（ART1）催化 Trim72 中精氨酸残基的单（ADP-核糖基）化，Trim72 是一种特异性表达在肌肉细胞中的蛋白质，参与细胞膜修复，其 ADP-核糖基化会增强。相比之下，ADP-核糖基化对上皮细胞中细胞膜修复的贡献仍不清楚。在这项研究中，我们研究了 ADP-核糖基化在上皮细胞的细胞膜修复中的作用。我们发现，使用链球菌溶血素-O 诱导细胞膜损伤后，多聚（ADP-核糖）聚合酶 1（PARP1）催化多聚（ADP-核糖基）化。在划痕实验中，使用非特异性 PARP 抑制剂 PJ34 或靶向 PARP1 的 shRNA 抑制 PARP1 活性会延迟伤口愈合，表明 PARP1 催化的多聚（ADP-核糖基）化在上皮细胞的细胞膜修复中发挥关键作用。
