Molecular and Cellular Biology Program, University of Washington, Seattle, Washington 98195.
G3 (Bethesda). 2013 Nov 6;3(11):1955-63. doi: 10.1534/g3.113.008250.
Eukaryotic origins of DNA replication undergo activation at various times in S-phase, allowing the genome to be duplicated in a temporally staggered fashion. In the budding yeast Saccharomyces cerevisiae, the activation times of individual origins are not intrinsic to those origins but are instead governed by surrounding sequences. Currently, there are two examples of DNA sequences that are known to advance origin activation time, centromeres and forkhead transcription factor binding sites. By combining deletion and linker scanning mutational analysis with two-dimensional gel electrophoresis to measure fork direction in the context of a two-origin plasmid, we have identified and characterized a 19- to 23-bp and a larger 584-bp DNA sequence that are capable of advancing origin activation time.
真核生物 DNA 复制的起始在 S 期的不同时间被激活，从而使基因组以时间交错的方式复制。在 budding 酵母 Saccharomyces cerevisiae 中，各个起始点的激活时间不是由起始点本身决定的，而是由周围的序列决定的。目前，有两个已知的 DNA 序列可以提前起始点的激活时间，分别是着丝粒和叉头转录因子结合位点。通过结合缺失和连接扫描突变分析以及二维凝胶电泳来测量在两个起始点质粒的情况下叉的方向，我们已经鉴定和描述了一个能够提前起始点激活时间的 19-23 个碱基对和一个更大的 584 个碱基对的 DNA 序列。
