Chen I-Peng, Mannuss Anja, Orel Nadiya, Heitzeberg Fabian, Puchta Holger
Botanical Institute II, University Karlsruhe, D-76128 Karlsruhe, Germany.
Plant Physiol. 2008 Apr;146(4):1786-96. doi: 10.1104/pp.108.116806. Epub 2008 Feb 29.
Rad5 is the key component in the Rad5-dependent error-free branch of postreplication repair in yeast (Saccharomyces cerevisiae). Rad5 is a member of the Snf2 ATPase/helicase family, possessing as a characteristic feature, a RING-finger domain embedded in the Snf2-helicase domain and a HIRAN domain. Yeast mutants are sensitive to DNA-damaging agents and reveal differences in homologous recombination. By sequence comparisons we were able to identify two homologs (AtRAD5a and AtRAD5b) in the Arabidopsis thaliana genome, sharing about 30% identity and 45% similarity to yeast Rad5. AtRad5a and AtRad5b have the same kind of domain organization with a higher degree of similarity to each other than to ScRad5. Surprisingly, both genes differ in function: whereas two independent mutants of Atrad5a are hypersensitive to the cross-linking agents mitomycin C and cis-platin and to a lesser extent to the methylating agent, methyl methane sulfonate, the Atrad5b mutants did not exhibit any sensitivity to all DNA-damaging agents tested. An Atrad5a/Atrad5b double mutant resembles the sensitivity phenotype of the Atrad5a single mutants. Moreover, in contrast to Atrad5b, the two Atrad5a mutants are deficient in homologous recombination after treatment with the double-strand break-inducing agent bleomycin. Our results suggest that the RAD5-dependent error-free branch of postreplication repair is conserved between yeast and plants, and that AtRad5a might be functionally homologous to ScRad5.
Rad5是酵母(酿酒酵母)复制后修复中依赖Rad5的无差错分支的关键组成部分。Rad5是Snf2 ATP酶/解旋酶家族的成员,其特征是在Snf2解旋酶结构域中嵌入一个RING指结构域和一个HIRAN结构域。酵母突变体对DNA损伤剂敏感,并在同源重组方面表现出差异。通过序列比较,我们能够在拟南芥基因组中鉴定出两个同源物(AtRAD5a和AtRAD5b),它们与酵母Rad5的同一性约为30%,相似性约为45%。AtRad5a和AtRad5b具有相同类型的结构域组织,彼此之间的相似程度高于与ScRad5的相似程度。令人惊讶的是,这两个基因在功能上有所不同:虽然Atrad5a的两个独立突变体对交联剂丝裂霉素C和顺铂高度敏感,对甲基化剂甲磺酸甲酯的敏感性较低,但Atrad5b突变体对所有测试的DNA损伤剂均未表现出任何敏感性。Atrad5a/Atrad5b双突变体类似于Atrad5a单突变体的敏感性表型。此外,与Atrad5b不同,两个Atrad5a突变体在用双链断裂诱导剂博来霉素处理后同源重组存在缺陷。我们的结果表明,复制后修复中依赖RAD5的无差错分支在酵母和植物之间是保守的,并且AtRad5a可能在功能上与ScRad5同源。