Interplay of macromolecular interactions during assembly of human DNA polymerase δ holoenzymes and initiation of DNA synthesis.
Interplay of macromolecular interactions during assembly of human DNA polymerase δ holoenzymes and initiation of DNA synthesis.
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人类 DNA 聚合酶 δ 全酶组装和 DNA 合成启动过程中大分子相互作用的相互作用。
DOI：
10.1101/2023.05.09.539896
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Hedglin,Mark
中科院分区：
文献类型：
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作者：
Norris,JessicaL;Rogers,LindseyO;Pytko,KaraG;Dannenberg,RachelL;Perreault,Samuel;Kaushik,Vikas;Kuppa,Sahiti;Antony,Edwin;Hedglin,Mark
在人类中，DNA聚合酶δ(POLδ)全酶由POLδ和过程性滑动钳、增殖细胞核抗原组成，在滞后链DNA复制、启动先导链DNA复制以及主要的DNA损伤修复和耐受途径中进行DNA合成。POLDNA全酶组装在引物/模板(P/T)连接处，在一个协调的过程中启动δ的合成，涉及主要的单链DNA结合蛋白复合体、复制蛋白A、加工性滑动钳夹加载器、复制因子C、增殖细胞核抗原和POLδ。这些因素中的每一个都与P/T结点产生独特的相互作用，并且最直接地相互作用。目前，这些大分子相互作用之间的相互作用在很大程度上是未知的。在本研究中，新颖的Förster共振能量转移(FRET)分析表明，在POLδ全酶的组装和DNA合成的启动过程中，RPA与P/T连接的动态相互作用保持在P/T连接，并容纳RFC、增殖细胞核抗原和POLδ，使每个过程的效率最大化。总的来说，这些研究极大地促进了我们对人类DNA复制和DNA修复的理解。
In humans, DNA polymerase δ (Pol δ) holoenzymes, comprised of Pol δ and the processivity sliding clamp, proliferating cell nuclear antigen (PCNA), carry out DNA synthesis during lagging strand DNA replication, initiation of leading strand DNA replication, and the major DNA damage repair and tolerance pathways. Pol δ holoenzymes are assembled at primer/template (P/T) junctions and initiate DNA synthesis in a coordinated process involving the major single strand DNA-binding protein complex, replication protein A (RPA), the processivity sliding clamp loader, replication factor C (RFC), PCNA, and Pol δ. Each of these factors interact uniquely with a P/T junction and most directly engage one another. Currently, the interplay between these macromolecular interactions is largely unknown. In the present study, novel Förster Resonance Energy Transfer (FRET) assays reveal that dynamic interactions of RPA with a P/T junction during assembly of a Pol δ holoenzyme and initiation of DNA synthesis maintain RPA at a P/T junction and accommodate RFC, PCNA, and Pol δ, maximizing the efficiency of each process. Collectively, these studies significantly advance our understanding of human DNA replication and DNA repair.