ELYS/MEL-28 chromatin association coordinates nuclear pore complex assembly and replication licensing.
ELYS/MEL-28 chromatin association coordinates nuclear pore complex assembly and replication licensing.
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DOI：
10.1016/j.cub.2007.08.041
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发表时间：
2007-10-09
期刊：
影响因子：
9.2
通讯作者：
Blow, J. Julian
中科院分区：
文献类型：
--
作者：
Gillespie, Peter J.;Khoudoli, Guennadi A.;Stewart, Graeme;Swedlow, Jason R.;Blow, J. Julian
非洲爪哇卵提取物在体外支持所有主要的细胞周期转变。我们使用蛋白质组学的方法来鉴定在体外细胞周期过程中染色质丰度发生变化的蛋白质。我们鉴定的蛋白质之一是Elys/MEL-28，它最近被描述为已知的核孔复合体(NPC)组装所需的最早作用因子。Elys与Nup107-160复合体相互作用，是其与染色质结合所必需的。ELYS包含一个AT-挂钩结构域，我们发现它与染色质有很高的亲和力。这个结构域可以与全长Elys竞争染色质结合，从而阻止NPC组装。这提供了Elys与染色质直接相互作用的证据，并且这种相互作用对于NPC的组装和细胞内染色体DNA的区隔是必不可少的。此外，我们检测到ELY和McM2-7复制许可蛋白之间在染色质上的物理关联。当McM2-7被阻止负载到染色质上时，Elys染色质负载、NPC组装和核生长都被延迟。由于在进入S阶段之前，核组装需要关闭许可，我们的结果表明这两个早期细胞周期事件相互协调的机制。
Xenopus egg extract supports all the major cell-cycle transitions in vitro. We have used a proteomics approach to identify proteins whose abundance on chromatin changes during the course of an in vitro cell cycle. One of the proteins we identified was ELYS/MEL-28, which has recently been described as the earliest-acting factor known to be required for nuclear pore complex (NPC) assembly. ELYS interacts with the Nup107-160 complex and is required for its association with chromatin. ELYS contains an AT-hook domain, which we show binds to chromatin with a high affinity. This domain can compete with full-length ELYS for chromatin association, thereby blocking NPC assembly. This provides evidence that ELYS interacts directly with chromatin and that this interaction is essential for NPC assembly and compartmentalization of chromosomal DNA within the cell. Furthermore, we detected a physical association on chromatin between ELYS and the Mcm2-7 replication-licensing proteins. ELYS chromatin loading, NPC assembly, and nuclear growth were delayed when Mcm2-7 was prevented from loading onto chromatin. Because nuclear assembly is required to shut down licensing prior to entry into S phase, our results suggest a mechanism by which these two early cell-cycle events are coordinated with one another.
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