Liu Z, Hossain G S, Islas-Osuna M A, Mitchell D L, Mount D W
Department of Molecular and Cellular Biology and Genetics Graduate Program, University of Arizona, Tucson, Arizona 85721, USA.
Plant J. 2000 Mar;21(6):519-28. doi: 10.1046/j.1365-313x.2000.00707.x.
To analyze plant mechanisms for resistance to UV radiation, mutants of Arabidopsis that are hypersensitive to UV radiation (designated uvh and uvr) have been isolated. UVR2 and UVR3 products were previously identified as photolyases that remove UV-induced pyrimidine dimers in the presence of visible light. Plants also remove dimers in the absence of light by an as yet unidentified dark repair mechanism and uvh1 mutants are defective in this mechanism. The UVH1 locus was mapped to chromosome 5 and the position of the UVH1 gene was further delineated by Agrobacterium-mediated transformation of the uvh1-1 mutant with cosmids from this location. Cosmid NC23 complemented the UV hypersensitive phenotype and restored dimer removal in the uvh1-1 mutant. The cosmid encodes a protein similar to the S. cerevisiae RAD1 and human XPF products, components of an endonuclease that excises dimers by nucleotide excision repair (NER). The uvh1-1 mutation creates a G to A transition in intron 5 of this gene, resulting in a new 3' splice site and introducing an in-frame termination codon. These results provide evidence that the Arabidopsis UVH1/AtRAD1 product is a subunit of a repair endonuclease. The previous discovery in Lilium longiflorum of a homolog of human ERCC1 protein that comprises the second subunit of the repair endonuclease provides additional evidence for the existence of the repair endonuclease in plants. The UVH1 gene is strongly expressed in flower tissue and also in other tissues, suggesting that the repair endonuclease is widely utilized for repair of DNA damage in plant tissues.
为了分析植物对紫外线辐射的抗性机制,已分离出对紫外线辐射高度敏感的拟南芥突变体(命名为uvh和uvr)。UVR2和UVR3产物先前被鉴定为光解酶,可在可见光存在下去除紫外线诱导的嘧啶二聚体。植物在无光条件下也可通过一种尚未明确的暗修复机制去除二聚体,而uvh1突变体在该机制中存在缺陷。UVH1基因座被定位到第5号染色体,通过用来自该位置的黏粒对uvh1 - 1突变体进行农杆菌介导的转化,进一步确定了UVH1基因的位置。黏粒NC23互补了紫外线敏感表型,并恢复了uvh1 - 1突变体中二聚体的去除。该黏粒编码一种与酿酒酵母RAD1和人类XPF产物相似的蛋白质,后者是一种通过核苷酸切除修复(NER)切除二聚体的核酸内切酶的组成成分。uvh1 - 1突变在该基因的第5内含子中产生了一个从G到A的转变,导致一个新的3'剪接位点,并引入了一个框内终止密码子。这些结果证明拟南芥UVH1/AtRAD1产物是修复核酸内切酶的一个亚基。先前在麝香百合中发现了人类ERCC1蛋白的同源物,它是修复核酸内切酶的第二个亚基,这为植物中存在修复核酸内切酶提供了额外证据。UVH1基因在花组织以及其他组织中强烈表达,表明修复核酸内切酶在植物组织中被广泛用于修复DNA损伤。