Lavrik O I, Khlimankov D Iu, Khodyreva S N
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division, Russian Academy of Sciences, Novosibirsk, 630090 Russia.
Mol Biol (Mosk). 2003 Jul-Aug;37(4):563-72.
Replication of eukaryotic DNA is driven by a protein complex, in which the central part is played by DNA polymerases. Synthesis with eukaryotic DNA polymerases alpha, delta, and epsilon involves various replication factors, including the replication protein A, replication factor C, proliferating cell nuclear antigen, etc. Replication enzymes and factors also participate in DNA repair, which is in an interplay with DNA replication. The function of the entire multicomponent system is regulated by protein--nucleic acid and protein--protein interactions. The eukaryotic replication complex was not isolated as a stable supramolecular structure, suggesting its dynamic organization. Hence X-ray analysis and other instrumental techniques are hardly suitable for studying this system. An alternative approach is affinity modification. Its most promising version involves in situ generation of photoreactive DNA replication intermediates. The review considers the recent progress in photoaffinity modification studies of DNA polymerases, eukaryotic replication factors, and their interactions with DNA replication intermediates.
真核生物DNA的复制由一种蛋白质复合体驱动,其中核心部分由DNA聚合酶发挥作用。真核生物DNA聚合酶α、δ和ε的合成涉及多种复制因子,包括复制蛋白A、复制因子C、增殖细胞核抗原等。复制酶和因子也参与DNA修复,其与DNA复制相互作用。整个多组分系统的功能由蛋白质-核酸和蛋白质-蛋白质相互作用调节。真核生物复制复合体并未作为稳定的超分子结构分离出来,这表明其具有动态组织。因此,X射线分析和其他仪器技术很难适用于研究该系统。一种替代方法是亲和修饰。其最有前景的形式涉及光反应性DNA复制中间体的原位生成。本文综述了DNA聚合酶、真核生物复制因子及其与DNA复制中间体相互作用的光亲和修饰研究的最新进展。