Novel method for site-specific induction of oxidative DNA damage reveals differences in recruitment of repair proteins to heterochromatin and euchromatin.
Novel method for site-specific induction of oxidative DNA damage reveals differences in recruitment of repair proteins to heterochromatin and euchromatin.
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DOI：
10.1093/nar/gkt1233
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发表时间：
2014-02
影响因子：
14.9
通讯作者：
Levine AS
中科院分区：
文献类型：
--
作者：
Lan L;Nakajima S;Wei L;Sun L;Hsieh CL;Sobol RW;Bruchez M;Van Houten B;Yasui A;Levine AS
活性氧（ROS）诱导的DNA损伤是通过碱基切除修复途径修复的。然而，染色质结构对BER蛋白募集到活细胞DNA损伤位点的影响尚不清楚。为了解决这个问题，我们开发了一种方法，通过将KillerRed（一种光刺激的ros诱导剂）与tet-阻遏因子（tetrr -KR）或转录激活因子（TA-KR）融合，特异性地产生ros诱导的DNA损伤。ttr - kr或TA-KR结合到U2OS细胞中特定基因组位点上的一个TRE盒（约90 kb）上，分别用于诱导异染色质或常染色质中的ROS损伤。我们发现DNA糖基酶在异染色质和常染色质中被有效地招募到DNA损伤中。与活性染色质相比，PARP1更有效地被招募到浓缩染色质内的DNA损伤中。相比之下，FEN1的募集在活性染色质内DNA损伤位点高度富集，以依赖于PCNA和转录激活的方式。这些结果表明，DNA氧化损伤在异染色质和常染色质中是不同的。
Reactive oxygen species (ROS)-induced DNA damage is repaired by the base excision repair pathway. However, the effect of chromatin structure on BER protein recruitment to DNA damage sites in living cells is poorly understood. To address this problem, we developed a method to specifically produce ROS-induced DNA damage by fusing KillerRed (KR), a light-stimulated ROS-inducer, to a tet-repressor (tetR-KR) or a transcription activator (TA-KR). TetR-KR or TA-KR, bound to a TRE cassette (∼90 kb) integrated at a defined genomic locus in U2OS cells, was used to induce ROS damage in hetero- or euchromatin, respectively. We found that DNA glycosylases were efficiently recruited to DNA damage in heterochromatin, as well as in euchromatin. PARP1 was recruited to DNA damage within condensed chromatin more efficiently than in active chromatin. In contrast, recruitment of FEN1 was highly enriched at sites of DNA damage within active chromatin in a PCNA- and transcription activation-dependent manner. These results indicate that oxidative DNA damage is differentially processed within hetero or euchromatin.
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影响因子：
64.5
作者：
Masui O;Bonnet I;Le Baccon P;Brito I;Pollex T;Murphy N;Hupé P;Barillot E;Belmont AS;Heard E
通讯作者：
Heard E
DOI：
10.1083/jcb.201205149
发表时间：
2012-12-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Menoni H;Hoeijmakers JH;Vermeulen W
通讯作者：
Vermeulen W
影响因子：
4.6
作者：
Barakat, Tahsin Stefan;Gribnau, Joost
通讯作者：
Gribnau, Joost
影响因子：
4.8
作者：
Pletnev, Sergei;Gurskaya, Nadya G.;Pletnev, Vladimir
通讯作者：
Pletnev, Vladimir
HMGB1：在碱基切除修复和相关功能中的作用。
DOI：
10.1016/j.bbagrm.2009.11.008
发表时间：
2010-01
影响因子：
4.7
作者：
Liu, Yuan;Prasad, Rajendra;Wilson, Samuel H.
通讯作者：
Wilson, Samuel H.