Schultz M C, Hockman D J, Harkness T A, Garinther W I, Altheim B A
Department of Biochemistry, University of Alberta, Edmonton, AB, Canada T6G 2H7.
Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):9034-9. doi: 10.1073/pnas.94.17.9034.
A simple in vitro system that supports chromatin assembly was developed for Saccharomyces cerevisiae. The assembly reaction is ATP-dependent, uses soluble histones and assembly factors, and generates physiologically spaced nucleosomes. We analyze the pathway of histone recruitment into nucleosomes, using this system in combination with genetic methods for the manipulation of yeast. This analysis supports the model of sequential recruitment of H3/H4 tetramers and H2A/H2B dimers into nucleosomes. Using a similar approach, we show that DNA ligase I can play an important role in template repair during assembly. These studies demonstrate the utility of this system for the combined biochemical and genetic analysis of chromatin assembly in yeast.
我们为酿酒酵母开发了一种支持染色质组装的简单体外系统。组装反应依赖ATP，使用可溶性组蛋白和组装因子，并生成具有生理间隔的核小体。我们结合用于操纵酵母的遗传方法，利用该系统分析组蛋白募集到核小体中的途径。该分析支持H3/H4四聚体和H2A/H2B二聚体依次募集到核小体中的模型。使用类似方法，我们表明DNA连接酶I在组装过程中的模板修复中可发挥重要作用。这些研究证明了该系统在酵母染色质组装的生化和遗传联合分析中的实用性。
