The Effect of the DNA Flanking the Lesion on Formation of the UvrB-DNA Preincision Complex

The Effect of the DNA Flanking the Lesion on Formation of the UvrB-DNA Preincision Complex
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病变侧翼 DNA 对 UvrB-DNA 预切割复合物形成的影响

DOI:
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发表时间:
2000
影响因子:
4.8
通讯作者:
N. Goosen
N. Goosen
中科院分区:
生物学2区
文献类型:
--
作者:
G. Moolenaar;V. Monaco;G. A. van der Marel;J. H. van Boom;R. Visse;N. Goosen

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UvrB-DNA切口前复合体在大肠杆菌的核苷酸切除修复中起关键作用。为了研究这种复合物的形成,构建了含有胆固醇加合物的DNA底物的衍生物。在加合物3 '侧的顶部或底部链上引入单链缺口,很可能通过释放DNA中的局部应力来稳定UvrB-DNA复合物。去除3 '切口处的两条DNA链仍然允许形成切口前复合体。然而,在损伤的5 '侧进行类似的修改,得到了不同的结果。在5 '切口处引入单链缺口完全消除了uvra介导的UvrB-DNA复合物的形成。两条DNA链在5 '切口处的缺失也阻止了UvrB通过uvra介导的加载到受损部位,但UvrB本身可以非常有效地结合。这表明,UvrB蛋白能够识别损伤,而不需要UvrA蛋白的媒人功能。我们的研究结果还表明,UvrB蛋白的uvra介导的负载是一个不对称的过程,从损伤的5 '侧开始。
The UvrB-DNA preincision complex plays a key role in nucleotide excision repair in Escherichia coli. To study the formation of this complex, derivatives of a DNA substrate containing a cholesterol adduct were constructed. Introduction of a single strand nick into either the top or the bottom strand at the 3′ side of the adduct stabilized the UvrB-DNA complex, most likely by the release of local stress in the DNA. Removal of both DNA strands up to the 3′ incision site still allowed formation of the preincision complex. Similar modifications at the 5′ side of the damage, however, gave different results. The introduction of a single strand nick at the 5′ incision site completely abolished the UvrA-mediated formation of the UvrB-DNA complex. Deletion of both DNA strands up to the 5′ incision site also prevented the UvrA-mediated loading of UvrB onto the damaged site, but UvrB by itself could bind very efficiently. This demonstrates that the UvrB protein is capable of recognizing damage without the matchmaker function of the UvrA protein. Our results also indicate that the UvrA-mediated loading of the UvrB protein is an asymmetric process, which starts at the 5′ side of the damage.
DOI: 10.1016/s0021-9258(18)45185-x
发表时间: 1987-09
期刊: The Journal of biological chemistry
影响因子: --
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DOI: --
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DOI: --
发表时间: 1989
期刊: The Journal of biological chemistry
影响因子: --
作者:
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