E1起始蛋白进行模板解链需要ATP依赖的TA碱基对小沟识别。
ATP-dependent minor groove recognition of TA base pairs is required for template melting by the E1 initiator protein.
作者信息
Schuck Stephen, Stenlund Arne
机构信息
Cold Spring Harbor Laboratory, 1 Bungtown Road, P.O. Cold Spring Harbor, NY 11724, USA.
出版信息
J Virol. 2007 Apr;81(7):3293-302. doi: 10.1128/JVI.02432-06. Epub 2007 Jan 3.
Abstract
Template melting is an essential step in the initiation of DNA replication, but the mechanism of template melting is unknown for any replicon. Here we demonstrate that melting of the bovine papillomavirus type 1 ori is a sequence-dependent process which relies on specific recognition of TA base pairs in the minor groove by the E1 initiator. We show that correct template melting is a prerequisite for the formation of a stable double hexamer with helicase activity and that ori mutants that fail to melt correctly are defective for ori unwinding and DNA replication in vivo. Our results also indicate that melting of the DNA is achieved by destabilization of the double helix along its length through multiple interactions with E1, each of which is responsible for melting of a few base pairs, resulting in the extensive melting that is required for initiation of DNA replication.
摘要
模板解链是DNA复制起始过程中的一个关键步骤，但对于任何复制子而言，模板解链的机制尚不清楚。在此，我们证明了1型牛乳头瘤病毒复制起点（ori）的解链是一个依赖序列的过程，该过程依赖于E1起始蛋白对小沟中TA碱基对的特异性识别。我们表明，正确的模板解链是形成具有解旋酶活性的稳定双六聚体的先决条件，并且不能正确解链的ori突变体在体内ori解旋和DNA复制方面存在缺陷。我们的结果还表明，DNA的解链是通过与E1的多重相互作用使双螺旋沿其长度方向不稳定而实现的，其中每一次相互作用都负责解开几个碱基对，从而导致DNA复制起始所需的广泛解链。
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