Structure of HsdS subunit from Thermoanaerobacter tengcongensis sheds lights on mechanism of dynamic opening and closing of type I methyltransferase.

Structure of HsdS subunit from Thermoanaerobacter tengcongensis sheds lights on mechanism of dynamic opening and closing of type I methyltransferase.
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腾康热厌氧杆菌的 HsdS 亚基结构揭示了 I 型甲基转移酶动态打开和关闭的机制

DOI:
10.1371/journal.pone.0017346
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发表时间:
2011-03-02
期刊:
影响因子:
3.7
通讯作者:
Liang D
Liang D
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Gao P;Tang Q;An X;Yan X;Liang D

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I型DNA甲基转移酶含有一个特异性亚基(HsdS)和两个修饰亚基(HsdM)。M. EcoKI-M2 S1甲基转移酶的电子显微镜模型显示了这种钳状酶的合理闭合状态,但开放状态的结构仍不清楚。腾冲嗜热厌氧菌特异性亚基(TTE-HsdS)的1.95 nm晶体结构显示了该亚基的未报道的开放式结构域间取向。基于TTE-HsdS的晶体结构和M. EcoKI-M2 S1的封闭态模型,我们构建了I型甲基转移酶的潜在开放态模型。突变研究表明TTE-HsdM亚基的两个α螺旋(aa 30 -59和aa 466 -495)是TTE-M2 S1复合物中重要的亚基间相互作用位点。DNA结合试验还强调了TTE-HsdM的C-末端区域对于TTE-M2 S1复合物结合DNA的重要性。在结构分析、生物化学实验和前人研究的基础上,我们提出了Ⅰ型甲基转移酶的一种动态开放和关闭机制。
Type I DNA methyltransferases contain one specificity subunit (HsdS) and two modification subunits (HsdM). The electron microscopy model of M.EcoKI-M2S1 methyltransferase shows a reasonable closed state of this clamp-like enzyme, but the structure of the open state is still unclear. The 1.95 Å crystal structure of the specificity subunit from Thermoanaerobacter tengcongensis (TTE-HsdS) shows an unreported open form inter-domain orientation of this subunit. Based on the crystal structure of TTE-HsdS and the closed state model of M.EcoKI-M2S1, we constructed a potential open state model of type I methyltransferase. Mutational studies indicated that two α-helices (aa30-59 and aa466-495) of the TTE-HsdM subunit are important inter-subunit interaction sites in the TTE-M2S1 complex. DNA binding assays also highlighted the importance of the C-terminal region of TTE-HsdM for DNA binding by the TTE-M2S1 complex. On the basis of structural analysis, biochemical experiments and previous studies, we propose a dynamic opening and closing mechanism for type I methyltransferase.
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