Domain swapping between FEN-1 and XPG defines regions in XPG that mediate nucleotide excision repair activity and substrate specificity.
Domain swapping between FEN-1 and XPG defines regions in XPG that mediate nucleotide excision repair activity and substrate specificity.
复制标题
FEN-1和XPG之间的域交换定义了XPG中介导核苷酸切除修复活性和底物特异性的区域。
DOI：
10.1093/nar/gkm092
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发表时间：
2007
影响因子：
14.9
通讯作者：
Schärer OD
中科院分区：
文献类型：
--
作者：
Hohl M;Dunand-Sauthier I;Staresincic L;Jaquier-Gubler P;Thorel F;Modesti M;Clarkson SG;Schärer OD
FEN-1和XPG是FEN-1结构特异性核酸酶家族的成员，它们共享一个保守的活性位点。FEN-1在DNA复制中起核心作用，而XPG则参与核苷酸切除修复（NER）。FEN-1和XPG都对叶片结构有活性，但只有XPG对气泡底物有活性。XPG的间隔区对于皮瓣底物上的核酸酶活性是必不可少的，但对于NER活性和气泡底物的有效处理是必需的。在这里，我们将XPG的间隔区插入FEN-1的核酸酶结构域之间，以测试该结构域是否足以赋予XPG样底物特异性和相关核酸酶上的NER活性。所得到的FEN-1-XPG杂交蛋白在皮瓣和气泡底物上都有活性，尽管水平较低。与FEN-1一样，FEN-1-XPG的活性受到含有1-nt 3 '瓣的双瓣底物的刺激，而XPG则没有这种底物偏好。虽然在体外没有检测到NER活性，但FEN-1-XPG杂交体在体内显示出大量的NER活性。因此，将XPG间隔区插入到FEN-1中会产生具有两种核酸酶生化特性的杂交蛋白，包括部分NER活性。
FEN-1 and XPG are members of the FEN-1 family of structure-specific nucleases, which share a conserved active site. FEN-1 plays a central role in DNA replication, whereas XPG is involved in nucleotide excision repair (NER). Both FEN-1 and XPG are active on flap structures, but only XPG cleaves bubble substrates. The spacer region of XPG is dispensable for nuclease activity on flap substrates but is required for NER activity and for efficient processing of bubble substrates. Here, we inserted the spacer region of XPG between the nuclease domains of FEN-1 to test whether this domain would be sufficient to confer XPG-like substrate specificity and NER activity on a related nuclease. The resulting FEN-1-XPG hybrid protein is active on flap and, albeit at low levels, on bubble substrates. Like FEN-1, the activity of FEN-1-XPG was stimulated by a double-flap substrate containing a 1-nt 3′ flap, whereas XPG does not show this substrate preference. Although no NER activity was detected in vitro, the FEN-1-XPG hybrid displays substantial NER activity in vivo. Hence, insertion of the XPG spacer region into FEN-1 results in a hybrid protein with biochemical properties reminiscent of both nucleases, including partial NER activity.
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影响因子：
3.9
作者：
Clarkson, SG
通讯作者：
Clarkson, SG
影响因子：
56.9
作者：
Houtsmuller, AB;Rademakers, S;Vermeulen, W
通讯作者：
Vermeulen, W
影响因子：
4.8
作者：
Kao, KI;Henricksen, LA;Bambara, RA
通讯作者：
Bambara, RA
影响因子：
4.8
作者：
Dunand-Sauthier, I;Hohl, M;Schärer, OD
通讯作者：
Schärer, OD
影响因子：
2.9
作者：
Kim, CY;Park, MS;Dyer, RB
通讯作者：
Dyer, RB