Physical and functional interaction between DDB and XPA in nucleotide excision repair.
Physical and functional interaction between DDB and XPA in nucleotide excision repair.
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DOI：
10.1093/nar/gkn964
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发表时间：
2009-02
影响因子：
14.9
通讯作者：
Matsunaga T
中科院分区：
文献类型：
--
作者：
Wakasugi M;Kasashima H;Fukase Y;Imura M;Imai R;Yamada S;Cleaver JE;Matsunaga T
损伤DNA结合蛋白(DDB)由DDB1和DDB2亚单位组成，可识别广泛的DNA损伤。在体外核苷酸切除修复(NER)反应中，DDB是必不可少的，但对环丁烷嘧啶二聚体(CPD)却有刺激作用。在这里，我们发现DDB与NER核心因子之一XPA直接相互作用，主要是通过DDB2亚单位，并且XPA中185-226个氨基酸残基对相互作用是重要的。有趣的是，先前在XP-A突变细胞系XP129中发现的导致从Arg-207替换为Gly的点突变，在体外和体内都降低了与DDB的相互作用。在一个包含R207G突变体XPA和其他核心NER因子的已确定的系统中，虽然突变体XPA能够进行基本的NER反应，但DDB未能刺激CPD的切除。此外，体内实验表明，突变体XPA被招募到DNA损伤部位的效率比野生型XPA低得多，并且不能支持在SV40转化的人细胞中通过异位表达DDB2来增强CPD修复。这些结果表明，DDB和XPA之间的物理相互作用在DDB介导的NER反应中起着重要作用。
Damaged DNA-binding protein (DDB), consisting of DDB1 and DDB2 subunits recognizes a wide spectrum of DNA lesions. DDB is dispensable for in vitro nucleotide excision repair (NER) reaction, but stimulates this reaction especially for cyclobutane pyrimidine dimer (CPD). Here we show that DDB directly interacts with XPA, one of core NER factors, mainly through DDB2 subunit and the amino-acid residues between 185 and 226 in XPA are important for the interaction. Interestingly, the point mutation causing the substitution from Arg-207 to Gly, which was previously identified in a XP-A revertant cell-line XP129, diminished the interaction with DDB in vitro and in vivo. In a defined system containing R207G mutant XPA and other core NER factors, DDB failed to stimulate the excision of CPD, although the mutant XPA was competent for the basal NER reaction. Moreover, in vivo experiments revealed that the mutant XPA is recruited to damaged DNA sites with much less efficiency compared with wild-type XPA and fails to support the enhancement of CPD repair by ectopic expression of DDB2 in SV40-transformed human cells. These results suggest that the physical interaction between DDB and XPA plays an important role in the DDB-mediated NER reaction.
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影响因子：
14.9
作者：
Buchko, GW;Ni, SS;Kennedy, MA
通讯作者：
Kennedy, MA
影响因子：
56.9
作者：
CHU, G;CHANG, E
通讯作者：
CHANG, E
DOI：
10.1073/pnas.0511160103
发表时间：
2006-02-21
影响因子：
11.1
作者：
Kapetanaki, MG;Guerrero-Santoro, J;Levine, AS
通讯作者：
Levine, AS
DOI：
10.1016/0921-8777(92)90049-9
发表时间：
1992-01-01
期刊：
MUTATION RESEARCH
影响因子：
--
作者：
KEENEY, S;WEIN, H;LINN, S
通讯作者：
LINN, S
影响因子：
14.9
作者：
Kobayashi, T;Takeuchi, S;Tanaka, K
通讯作者：
Tanaka, K