Cleavage of phosphorothioated DNA and methylated DNA by the type IV restriction endonuclease ScoMcrA.
Cleavage of phosphorothioated DNA and methylated DNA by the type IV restriction endonuclease ScoMcrA.
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IV 型限制性内切酶 ScoMcrA 对硫代磷酸化 DNA 和甲基化 DNA 的切割
DOI：
10.1371/journal.pgen.1001253
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发表时间：
2010-12-23
期刊：
影响因子：
4.5
通讯作者：
He X
中科院分区：
文献类型：
--
作者：
Liu G;Ou HY;Wang T;Li L;Tan H;Zhou X;Rajakumar K;Deng Z;He X
许多分类学上不同的原核生物通过在特定序列处用硫原子替换非桥接氧来酶促修饰它们的DNA。这种DNA S-修饰（硫代磷酸化）的生物学意义尚不清楚。我们观察到，同时表达来自变铅青链霉菌66的dndA-E基因簇（负责DNA S-修饰）和推定的天蓝色链霉菌A（3）2 IV型甲基依赖性限制性内切酶ScoA 3 McrA（Sco 4631）导致同一宿主中的细胞死亡。ScoA 3 McrA的His标记的衍生物在各自的修饰位点附近在体外切割S修饰的DNA和Dcm甲基化的DNA。双链切割发生在距离硫代磷酸酯连接16-28个核苷酸处。DNA酶I足迹法证明了ScoA 3 McrA与Dcm甲基化位点的结合，但在切割条件下，在S-修饰位点没有检测到明显的结合。这是McrA同源物体外核酸内切酶活性的首次报道，也是特异性切割S修饰DNA的酶的首次证明。
Many taxonomically diverse prokaryotes enzymatically modify their DNA by replacing a non-bridging oxygen with a sulfur atom at specific sequences. The biological implications of this DNA S-modification (phosphorothioation) were unknown. We observed that simultaneous expression of the dndA-E gene cluster from Streptomyces lividans 66, which is responsible for the DNA S-modification, and the putative Streptomyces coelicolor A(3)2 Type IV methyl-dependent restriction endonuclease ScoA3McrA (Sco4631) leads to cell death in the same host. A His-tagged derivative of ScoA3McrA cleaved S-modified DNA and also Dcm-methylated DNA in vitro near the respective modification sites. Double-strand cleavage occurred 16–28 nucleotides away from the phosphorothioate links. DNase I footprinting demonstrated binding of ScoA3McrA to the Dcm methylation site, but no clear binding could be detected at the S-modified site under cleavage conditions. This is the first report of in vitro endonuclease activity of a McrA homologue and also the first demonstration of an enzyme that specifically cleaves S-modified DNA.
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影响因子：
12.3
作者：
Fukuda, Eri;Kaminska, Katarzyna H.;Bujnicki, Janusz M.;Kobayashi, Ichizo
通讯作者：
Kobayashi, Ichizo
影响因子：
2.1
作者：
Flett, F;Mersinias, V;Smith, CP
通讯作者：
Smith, CP
影响因子：
14.9
作者：
Dalgaard, JZ;Klar, AJ;Mian, IS
通讯作者：
Mian, IS
影响因子：
14.9
作者：
JANULAITIS, A;PETRUSYTE, M;BUTKUS, V
通讯作者：
BUTKUS, V
影响因子：
2.1
作者：
Gonzalez-Ceron, Gabriela;Josue Miranda-Olivares, Osvaldo;Servin-Gonzalez, Luis
通讯作者：
Servin-Gonzalez, Luis