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首页> 外文期刊>Journal of bacteriology >UvrD2 Is Essential in Mycobacterium tuberculosis, but Its Helicase Activity Is Not Required
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UvrD2 Is Essential in Mycobacterium tuberculosis, but Its Helicase Activity Is Not Required

机译：UvrD2在结核分枝杆菌中必不可少，但不需要其解旋酶活性

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UvrD is an SF1 family helicase involved in DNA repair that is widely conserved in bacteria. Mycobacterium tuberculosishas two annotated UvrD homologues; here we investigate the role of UvrD2. The uvrD2gene at its native locus could be knocked out only in the presence of a second copy of the gene, demonstrating that uvrD2is essential. Analysis of the putative protein domain structure of UvrD2 shows a distinctive domain architecture, with an extended C terminus containing an HRDC domain normally found in SF2 family helicases and a linking domain carrying a tetracysteine motif. Truncated constructs lacking the C-terminal domains of UvrD2 were able to compensate for the loss of the chromosomal copy, showing that these C-terminal domains are not essential. Although UvrD2 is a functional helicase, a mutant form of the protein lacking helicase activity was able to permit deletion of uvrD2at its native locus. However, a mutant protein unable to hydrolyze ATP or translocate along DNA was not able to compensate for lack of the wild-type protein. Therefore, we concluded that the essential role played by UvrD2 is unlikely to involve its DNA unwinding activity and is more likely to involve DNA translocation and, possibly, protein displacement.
机译：UvrD是涉及DNA修复的SF1家族解旋酶，在细菌中广泛保守。结核分枝杆菌有两个带注释的UvrD同源物;在这里，我们研究UvrD2的作用。仅在该基因的第二个拷贝存在时，才能敲除其天然位点的uvrD2基因，这表明uvrD2是必不可少的。对UvrD2的推定蛋白质结构域结构的分析显示出独特的结构域结构，其延伸的C末端包含通常在SF2家族解旋酶中发现的HRDC结构域和带有四半胱氨酸基序的连接结构域。缺少UvrD2 C末端结构域的截短构建体能够补偿染色体复制的丢失，表明这些C末端结构域不是必需的。尽管UvrD2是一种功能性解旋酶，但缺少解旋酶活性的蛋白质突变形式却能够使uvrD2在其天然位点缺失。但是，无法水解ATP或沿DNA转运的突变蛋白无法补偿野生型蛋白的缺乏。因此，我们得出的结论是，UvrD2发挥的基本作用不太可能涉及其DNA解链活性，而更可能涉及DNA易位以及可能的蛋白质置换。

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