Cryo-EM Structural Biology Laboratory, Van Andel Research Institute, Grand Rapids, MI, USA.
Biochemistry and Structural Biology Graduate Program, Stony Brook University, Stony Brook, NY, USA.
Adv Exp Med Biol. 2017;1042:207-228. doi: 10.1007/978-981-10-6955-0_10.
Eukaryotic replication proteins are highly conserved, and thus study of Saccharomyces cerevisiae replication can inform about this central process in higher eukaryotes including humans. The S. cerevisiae replisome is a large and dynamic assembly comprised of ~50 proteins. The core of the replisome is composed of 31 different proteins including the 11-subunit CMG helicase; RFC clamp loader pentamer; PCNA clamp; the heteroligomeric DNA polymerases ε, δ, and α-primase; and the RPA heterotrimeric single strand binding protein. Many additional protein factors either travel with or transiently associate with these replisome proteins at particular times during replication. In this chapter, we summarize several recent structural studies on the S. cerevisiae replisome and its subassemblies using single particle electron microscopy and X-ray crystallography. These recent structural studies have outlined the overall architecture of a core replisome subassembly and shed new light on the mechanism of eukaryotic replication.
真核复制蛋白高度保守，因此对酿酒酵母（Saccharomyces cerevisiae）复制的研究可以为包括人类在内的高等真核生物的这一核心过程提供信息。酿酒酵母的复制体是一个由~50 种蛋白质组成的大型动态组装体。复制体的核心由 31 种不同的蛋白质组成，包括 11 亚基的 CMG 解旋酶；RFC 夹装载五聚体；PCNA 夹；异源三聚体 DNA 聚合酶 ε、δ 和 α-引发酶；和 RPA 异源三聚体单链结合蛋白。许多其他的蛋白质因子要么在复制过程中的特定时间与这些复制体蛋白一起移动，要么与之短暂结合。在本章中，我们使用单颗粒电子显微镜和 X 射线晶体学对酿酒酵母复制体及其亚基进行了几项最近的结构研究进行了总结。这些最近的结构研究概述了核心复制体亚基的整体结构，并为真核复制的机制提供了新的见解。
