Doucet-Chabeaud G, Godon C, Brutesco C, de Murcia G, Kazmaier M
CEA/Cadarache, DSV-DEVM-Laboratoire de Radiobiologie Végétale, Saint Paul-lez-Durance, France.
Mol Genet Genomics. 2001 Aug;265(6):954-63. doi: 10.1007/s004380100506.
By screening for Arabidopsis genes activated by ionising radiation (IR)-induced DNA damage, we have isolated a cDNA hybridising with a 3.2-kb mRNA that accumulates rapidly and strongly in irradiated cell suspensions or whole plants. The cDNA codes for a 110-kDa protein that is highly homologous to the 116-kDa vertebrate poly(ADP-ribose) polymerase (PARP-1). It is recognised by a human anti-PARP-1 antibody, binds efficiently to DNA strand interruptions in vitro, and catalyses DNA damage-dependent (ADP-ribose) polymer synthesis. We have named this protein AtPARP-1. We have also extended our observations to the Arabidopsis app (AtPARP-2) gene, demonstrating for the first time that IR-induced DNA strand interruptions induce rapid and massive accumulation of AtPARP-1 and AtPARP-2 transcripts, whereas dehydration and cadmium preferentially induce the accumulation of AtPARP-2 transcripts. The IR-induced PARP gene expression seen in Arabidopsis is in striking contrast to the post-translational activation of the PARP-1 protein that is associated with genotoxic stress in animal cells. AtPARP-1 transcripts accumulate in all plant organs after exposure to ionising radiation, but this is followed by an increase in AtPARP-1 protein levels only in tissues that contain large amounts of actively dividing cells. This cell-type specific accumulation of AtPARP-1 protein in response to DNA damage is compatible with a role for the AtPARP-1 protein in the maintenance of DNA integrity during replication, similar to the role of "guardian of the genome" attributed to its animal counterpart.
通过筛选拟南芥中由电离辐射(IR)诱导的DNA损伤所激活的基因,我们分离出了一个与3.2 kb mRNA杂交的cDNA,该mRNA在受辐照的细胞悬浮液或整株植物中迅速且大量积累。该cDNA编码一种110 kDa的蛋白质,它与116 kDa的脊椎动物聚(ADP - 核糖)聚合酶(PARP - 1)高度同源。它能被人抗PARP - 1抗体识别,在体外能有效结合DNA链断裂处,并催化依赖DNA损伤的(ADP - 核糖)聚合物合成。我们将这种蛋白质命名为AtPARP - 1。我们还将观察扩展到了拟南芥app(AtPARP - 2)基因,首次证明IR诱导的DNA链断裂会诱导AtPARP - 1和AtPARP - 2转录本迅速大量积累,而脱水和镉则优先诱导AtPARP - 2转录本的积累。拟南芥中IR诱导的PARP基因表达与动物细胞中与遗传毒性应激相关的PARP - 1蛋白的翻译后激活形成了鲜明对比。AtPARP - 1转录本在暴露于电离辐射后在所有植物器官中积累,但随后仅在含有大量活跃分裂细胞的组织中AtPARP - 1蛋白水平增加。这种AtPARP - 1蛋白响应DNA损伤的细胞类型特异性积累与AtPARP - 1蛋白在复制过程中维持DNA完整性的作用相一致,类似于赋予其动物对应物的“基因组守护者”的作用。