早期启动与核周锚定的复制原点的连接。
Early initiation of a replication origin tethered at the nuclear periphery.
机构信息
Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
出版信息
J Cell Sci. 2010 Apr 1;123(Pt 7):1015-9. doi: 10.1242/jcs.060392. Epub 2010 Mar 2.
Abstract
Peripheral nuclear localization of chromosomal loci correlates with late replication in yeast and metazoan cells. To test whether peripheral positioning can impose late replication, we examined whether artificial tethering of an early-initiating replication origin to the nuclear periphery delays its replication in budding yeast. We tested the effects of three different peripheral tethering constructs on the time of replication of the early replication origin ARS607. Using the dense-isotope transfer method to assess replication time, we found that ARS607 still replicates early when tethered to the nuclear periphery using the Yif1 protein or a fragment of Sir4, whereas tethering using a Yku80 construct produces only a very slight replication delay. Single-cell microscopic analysis revealed no correlation between peripheral positioning of ARS607 in individual cells and delayed replication. Overall, our results demonstrate that a replication origin can initiate replication early in S phase, even if artificially relocated to the nuclear periphery.
摘要
染色体位置的核周定位与酵母和后生动物细胞的晚期复制相关。为了测试外周定位是否可以施加晚期复制，我们研究了将早期起始复制原点人为地固定在核周是否会延迟其在芽殖酵母中的复制。我们测试了三种不同的外周固定构建体对早期复制原点 ARS607 复制时间的影响。使用密集同位素转移方法评估复制时间，我们发现当使用 Yif1 蛋白或 Sir4 的片段将 ARS607 固定到核周时，它仍然在早期复制，而使用 Yku80 构建体固定时仅产生非常轻微的复制延迟。单细胞显微镜分析显示，ARS607 在单个细胞中的外周定位与延迟复制之间没有相关性。总体而言，我们的结果表明，即使人为地将复制原点重新定位到核周，它也可以在 S 期早期开始复制。
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