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拟南芥DNA连接酶IV受γ射线诱导,并与双链断裂修复蛋白XRCC4的拟南芥同源物相互作用。

Arabidopsis DNA ligase IV is induced by gamma-irradiation and interacts with an Arabidopsis homologue of the double strand break repair protein XRCC4.

作者信息

West C E, Waterworth W M, Jiang Q, Bray C M

机构信息

School of Biological Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK.

出版信息

Plant J. 2000 Oct;24(1):67-78. doi: 10.1046/j.1365-313x.2000.00856.x.

Abstract

Rejoining of single- and double-strand breaks (DSBs) introduced in DNA during replication, recombination, and DNA damage is catalysed by DNA ligase enzymes. Eukaryotes possess multiple DNA ligase enzymes, each having distinct roles in cellular metabolism. Double-strand breaks in DNA, which can occur spontaneously in the cell or be induced experimentally by gamma-irradiation, represent one of the most serious threats to genomic integrity. Non-homologous end joining (NHEJ) rather than homologous recombination is the major pathway for repair of DSBs in organisms with complex genomes, including humans and plants. DNA ligase IV in Saccharomyces cerevisiae and humans catalyses the final step in the NHEJ pathway of DSB repair. In this study we identify an Arabidopsis thaliana homologue (AtLIG4) of human and S. cerevisiae DNA ligase IV which is shown to encode an ATP-dependent DNA ligase with a theoretical molecular mass of 138 kDa and 48% similarity in amino-acid sequence to the human DNA ligase IV. Yeast two-hybrid analysis demonstrated a strong interaction between A. thaliana DNA ligase IV and the A. thaliana homologue of the human DNA ligase IV-binding protein XRCC4. This interaction is shown to be mediated via the tandem BRCA C-terminal domains of A. thaliana DNA ligase IV protein. Expression of AtLIG4 is induced by gamma-irradiation but not by UVB irradiation, consistent with an in vivo role for the A. thaliana DNA ligase IV in DSB repair.

摘要

在复制、重组及DNA损伤过程中引入到DNA中的单链和双链断裂(DSB)的重新连接由DNA连接酶催化。真核生物拥有多种DNA连接酶,每种在细胞代谢中都有不同作用。DNA双链断裂可在细胞中自发发生,也可通过γ射线实验诱导产生,它是对基因组完整性最严重的威胁之一。在包括人类和植物在内的具有复杂基因组的生物体中,非同源末端连接(NHEJ)而非同源重组是DSB修复的主要途径。酿酒酵母和人类中的DNA连接酶IV催化DSB修复的NHEJ途径中的最后一步。在本研究中,我们鉴定出了人类和酿酒酵母DNA连接酶IV的拟南芥同源物(AtLIG4),它被证明编码一种依赖ATP的DNA连接酶,理论分子量为138 kDa,氨基酸序列与人类DNA连接酶IV有48%的相似性。酵母双杂交分析表明拟南芥DNA连接酶IV与人类DNA连接酶IV结合蛋白XRCC4的拟南芥同源物之间有强烈相互作用。这种相互作用被证明是通过拟南芥DNA连接酶IV蛋白的串联BRCA C末端结构域介导的。AtLIG4的表达由γ射线诱导,但不由UVB辐射诱导,这与拟南芥DNA连接酶IV在体内DSB修复中的作用一致。

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