Novel inhibition of archaeal family-D DNA polymerase by uracil.

Novel inhibition of archaeal family-D DNA polymerase by uracil.
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DOI:
10.1093/nar/gkt083
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发表时间:
2013-04
影响因子:
14.9
通讯作者:
Henneke G
Henneke G
中科院分区:
生物学2区
文献类型:
--
作者:
Richardson TT;Gilroy L;Ishino Y;Connolly BA;Henneke G

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古细菌家族d DNA聚合酶被DNA模板链中尿嘧啶的存在所抑制。当酶遇到尿嘧啶时,以下三个参数发生变化:DNA结合增加约2倍,聚合速度减慢约5倍,3 ‘ -5 ’校对外切酶活性受到约2倍的刺激。这些变化共同导致聚合活性的显著降低和净DNA合成的减少。Pol D似乎与模板链尿嘧啶相互作用,与它在复制叉前面的距离无关。聚合并不停止在一个特定的位置相对于尿嘧啶,而是普遍减少DNA合成被观察到。“反式”抑制,尿嘧啶在DNA链上不被复制的Pol D减慢也被观察到。有人提出,Pol D能够通过环出单链模板与尿嘧啶相互作用,允许碱基和引物-模板连接处同时接触,从而产生延伸能力减弱的聚合酶- dna复合物。
Archaeal family-D DNA polymerase is inhibited by the presence of uracil in DNA template strands. When the enzyme encounters uracil, following three parameters change: DNA binding increases roughly 2-fold, the rate of polymerization slows by a factor of ∼5 and 3′–5′ proof-reading exonuclease activity is stimulated by a factor of ∼2. Together these changes result in a significant decrease in polymerization activity and a reduction in net DNA synthesis. Pol D appears to interact with template strand uracil irrespective of its distance ahead of the replication fork. Polymerization does not stop at a defined location relative to uracil, rather a general decrease in DNA synthesis is observed. ‘Trans’ inhibition, the slowing of Pol D by uracil on a DNA strand not being replicated is also observed. It is proposed that Pol D is able to interact with uracil by looping out the single-stranded template, allowing simultaneous contact of both the base and the primer-template junction to give a polymerase-DNA complex with diminished extension ability.
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