Haracska Lajos, Acharya Narottam, Unk Ildiko, Johnson Robert E, Hurwitz Jerard, Prakash Louise, Prakash Satya
Sealy Center for Molecular Science, University of Texas Medical Branch, 6.104 Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555-1061, USA.
Mol Cell Biol. 2005 Feb;25(3):1183-90. doi: 10.1128/MCB.25.3.1183-1190.2005.
DNA polymerases (Pols) of the Y family rescue stalled replication forks by promoting replication through DNA lesions. Humans have four Y family Pols, eta, iota, kappa, and Rev1, of which Pols eta, iota, and kappa have been shown to physically interact with proliferating cell nuclear antigen (PCNA) and be functionally stimulated by it. However, in sharp contrast to the large increase in processivity that PCNA binding imparts to the replicative Pol, Poldelta, the processivity of Y family Pols is not enhanced upon PCNA binding. Instead, PCNA binding improves the efficiency of nucleotide incorporation via a reduction in the apparent K(m) for the nucleotide. Here we show that Poliota interacts with PCNA via only one of its conserved PCNA binding motifs, regardless of whether PCNA is bound to DNA or not. The mode of PCNA binding by Poliota is quite unlike that in Poldelta, where multisite interactions with PCNA provide for a very tight binding of the replicating Pol with PCNA. We discuss the implications of these observations for the accuracy of DNA synthesis during translesion synthesis and for the process of Pol exchange at the lesion site.
Y家族的DNA聚合酶(Pols)通过促进DNA损伤处的复制来拯救停滞的复制叉。人类有四种Y家族的Pols,即eta、iota、kappa和Rev1,其中Pols eta、iota和kappa已被证明能与增殖细胞核抗原(PCNA)发生物理相互作用,并受到其功能刺激。然而,与PCNA结合赋予复制性Pol δ大幅提高的持续合成能力形成鲜明对比的是,Y家族Pols的持续合成能力在PCNA结合后并未增强。相反,PCNA结合通过降低核苷酸的表观K(m)来提高核苷酸掺入的效率。在这里我们表明,无论PCNA是否与DNA结合,Poliota仅通过其一个保守的PCNA结合基序与PCNA相互作用。Poliota与PCNA的结合模式与Pol δ截然不同,在Pol δ中,与PCNA的多位点相互作用使得复制性Pol与PCNA紧密结合。我们讨论了这些观察结果对跨损伤合成过程中DNA合成准确性以及损伤位点处Pol交换过程的影响。