The unstructured C-terminal extension of UvrD interacts with UvrB, but is dispensable for nucleotide excision repair.
The unstructured C-terminal extension of UvrD interacts with UvrB, but is dispensable for nucleotide excision repair.
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DOI：
10.1016/j.dnarep.2009.08.005
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发表时间：
2009-11-02
期刊：
影响因子：
3.8
通讯作者：
Savery NJ
中科院分区：
文献类型：
--
作者：
Manelyte L;Guy CP;Smith RM;Dillingham MS;McGlynn P;Savery NJ
在细菌的核苷酸切除修复（NER）过程中，UvrC核酸酶和含有DNA损伤的短寡核苷酸被超家族1A解旋酶UvrD从切口后复合体中去除。解旋酶经常通过与伴侣蛋白的相互作用来调节，免疫沉淀实验先前表明，UvrD与UvrB相互作用，UvrB是切口后复合体的一个组成部分。我们使用2-杂化分析和表面等离子体共振光谱分析了这种相互作用，发现UvrD的n端结构域和c端非结构化区域与UvrB相互作用。我们分析了缺乏非结构化c端区域的截断的UvrD蛋白的特性，发现它对单链DNA的亲和力降低，但保留了从切口后核苷酸切除修复复合体中置换UvrC和含有病变的寡核苷酸的能力。因此，UvrD的c端区域与UvrB的相互作用并不是UvrD在NER期间分解切口后复合体的机制的基本特征。在进一步的实验中，我们发现来自嗜热脂肪芽孢杆菌的PcrA解螺旋酶也可以取代切口后NER复合体中的UvrC和切除的寡核苷酸，这支持了PcrA在革兰氏阳性生物NER过程中发挥类似uvrd功能的观点。
During nucleotide excision repair (NER) in bacteria the UvrC nuclease and the short oligonucleotide that contains the DNA lesion are removed from the post-incision complex by UvrD, a superfamily 1A helicase. Helicases are frequently regulated by interactions with partner proteins, and immunoprecipitation experiments have previously indicated that UvrD interacts with UvrB, a component of the post-incision complex. We examined this interaction using 2-hybrid analysis and surface plasmon resonance spectroscopy, and found that the N-terminal domain and the unstructured region at the C-terminus of UvrD interact with UvrB. We analysed the properties of a truncated UvrD protein that lacked the unstructured C-terminal region and found that it showed a diminished affinity for single-stranded DNA, but retained the ability to displace both UvrC and the lesion-containing oligonucleotide from a post-incision nucleotide excision repair complex. The interaction of the C-terminal region of UvrD with UvrB is therefore not an essential feature of the mechanism by which UvrD disassembles the post-incision complex during NER. In further experiments we showed that PcrA helicase from Bacillus stearothermophilus can also displace UvrC and the excised oligonucleotide from a post-incision NER complex, which supports the idea that PcrA performs a UvrD-like function during NER in Gram-positive organisms.
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DOI：
10.1073/pnas.96.21.11717
发表时间：
1999-10-12
影响因子：
11.1
作者：
Machius, M;Henry, L;Deisenhofer, J
通讯作者：
Deisenhofer, J
DOI：
10.1073/pnas.82.15.4925
发表时间：
1985-01-01
影响因子：
11.1
作者：
CARON, PR;KUSHNER, SR;GROSSMAN, L
通讯作者：
GROSSMAN, L
影响因子：
3.2
作者：
Mechanic, LE;Latta, NE;Matson, SW
通讯作者：
Matson, SW
大肠杆菌K-12的构造框架，单基因敲除突变体：Keio Collection。
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
14.9
作者：
Kirsch, RD;Joly, E
通讯作者：
Joly, E