van Attikum Haico, Bundock Paul, Overmeer René M, Lee Lan-Ying, Gelvin Stanton B, Hooykaas Paul J J
Institute of Biology, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands.
Nucleic Acids Res. 2003 Jul 15;31(14):4247-55. doi: 10.1093/nar/gkg458.
The joining of breaks in the chromosomal DNA backbone by ligases in processes of replication, recombination and repair plays a crucial role in the maintenance of genomic stability. Four ATP-dependent ligases, designated DNA ligases I-IV, have been identified in higher eukaryotes, and each one has distinct functions. In mammals and yeast, DNA ligase IV is exclusively involved in the repair of DNA double-strand breaks by non-homologous end joining. Recently, an Arabidopsis thaliana orthologue of the yeast and mammalian DNA ligase IV gene was found and termed AtLIG4. Here we describe the isolation and functional characterisation of a plant line with a T-DNA insertion in the AtLIG4 gene. Plants homozygous for the T-DNA insertion did not display any growth or developmental defects and were fertile. However, mutant seedlings were hypersensitive to the DNA-damaging agents methyl methanesulfonate and X-rays, demonstrating that AtLIG4 is required for the repair of DNA damage. Recently, we showed that a yeast lig4 mutant is deficient in Agrobacterium T-DNA integration. However, using tumorigenesis and germline transformation assays, we found that the plant AtLIG4 mutant is not impaired in T-DNA integration. Thus, in contrast to yeast, DNA ligase IV is not required for T-DNA integration in plants.
在复制、重组和修复过程中,连接酶将染色体DNA主链中的断裂处连接起来,这在维持基因组稳定性方面起着至关重要的作用。在高等真核生物中已鉴定出四种依赖ATP的连接酶,分别命名为DNA连接酶I-IV,且每种连接酶都有不同的功能。在哺乳动物和酵母中,DNA连接酶IV专门参与通过非同源末端连接修复DNA双链断裂。最近,发现了酵母和哺乳动物DNA连接酶IV基因的拟南芥同源物,并将其命名为AtLIG4。在此,我们描述了一个AtLIG4基因中存在T-DNA插入的植物株系的分离和功能特性。T-DNA插入纯合的植物没有表现出任何生长或发育缺陷,并且可育。然而,突变幼苗对DNA损伤剂甲磺酸甲酯和X射线高度敏感,表明AtLIG4是修复DNA损伤所必需的。最近,我们表明酵母lig4突变体在农杆菌T-DNA整合方面存在缺陷。然而,通过肿瘤发生和种系转化试验,我们发现植物AtLIG4突变体在T-DNA整合方面没有受损。因此,与酵母不同,植物中的T-DNA整合不需要DNA连接酶IV。