An archaeal family-B DNA polymerase variant able to replicate past DNA damage: occurrence of replicative and translesion synthesis polymerases within the B family.
An archaeal family-B DNA polymerase variant able to replicate past DNA damage: occurrence of replicative and translesion synthesis polymerases within the B family.
复制标题
DOI：
10.1093/nar/gku683
复制
发表时间：
2014-09
影响因子：
14.9
通讯作者：
Connolly BA
中科院分区：
文献类型：
--
作者：
Jozwiakowski SK;Keith BJ;Gilroy L;Doherty AJ;Connolly BA
描述了一种来自古细菌Thermococcus gorgonarius（Tgo-Pol）的高保真B家族DNA聚合酶的突变体，该突变体能够复制过去的DNA损伤。功能的获得需要将Tgo-Pol的指状结构域中的三个氨基酸环区替换为天然存在于真核Pol β（一种B家族跨损伤合成聚合酶）中的较长形式。3′-5′校正核酸外切酶活性的失活也是必要的。由此产生的Tgo-Pol Z1变体擅长从碱基错配开始复制，并且可以读取受损的碱基，例如脱碱基位点和胸腺嘧啶光二聚体。Tgo-Pol Z1也擅长从终止相对异常碱基的引物延伸。Tgo-Pol Zl的保真度降低，具有在G：C碱基对处进行改变的显著趋势。总之，这些结果表明，手指结构域的环区可能发挥关键作用，在确定是否家族B酶福尔斯属于准确的基因组复制的类别，或者是一个容易出错的translesion合成聚合酶。Tgo-Pol Z1也可用于扩增受损的DNA。
A mutant of the high fidelity family-B DNA polymerase from the archaeon Thermococcus gorgonarius (Tgo-Pol), able to replicate past DNA lesions, is described. Gain of function requires replacement of the three amino acid loop region in the fingers domain of Tgo-Pol with a longer version, found naturally in eukaryotic Pol ζ (a family-B translesion synthesis polymerase). Inactivation of the 3′–5′ proof-reading exonuclease activity is also necessary. The resulting Tgo-Pol Z1 variant is proficient at initiating replication from base mismatches and can read through damaged bases, such as abasic sites and thymine photo-dimers. Tgo-Pol Z1 is also proficient at extending from primers that terminate opposite aberrant bases. The fidelity of Tgo-Pol Z1 is reduced, with a marked tendency to make changes at G:C base pairs. Together, these results suggest that the loop region of the fingers domain may play a critical role in determining whether a family-B enzyme falls into the accurate genome-replicating category or is an error-prone translesion synthesis polymerase. Tgo-Pol Z1 may also be useful for amplification of damaged DNA.
登录
查看更多内容
影响因子：
64.5
作者：
Franklin, MC;Wang, JM;Steitz, TA
通讯作者：
Steitz, TA
影响因子：
5.6
作者：
Firbank, Susan J.;Wardle, Josephine;Connolly, Bernard A.
通讯作者：
Connolly, Bernard A.
影响因子：
14.9
作者：
BRAITHWAITE, DK;ITO, J
通讯作者：
ITO, J
影响因子：
5.3
作者：
Johnson, RE;Haracska, L;Prakash, L
通讯作者：
Prakash, L
影响因子：
16.8
作者：
Hogg, Matthew;Osterman, Pia;Johansson, Erik
通讯作者：
Johansson, Erik