Biphasic chromatin binding of histone chaperone FACT during eukaryotic chromatin DNA replication.
Biphasic chromatin binding of histone chaperone FACT during eukaryotic chromatin DNA replication.
复制标题
真核染色质 DNA 复制过程中组蛋白伴侣 FACT 的双相染色质结合。
DOI：
10.1016/j.bbamcr.2011.01.002
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发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Tada S
中科院分区：
文献类型：
--
作者：
Kundu LR;Seki M;Watanabe N;Murofushi H;Furukohri A;Waga S;Score AJ;Blow JJ;Horikoshi M;Enomoto T;Tada S
促进染色质转录（FACT）复合物影响染色质背景下的核DNA交易。虽然FACT参与真核生物DNA复制的机制已经被揭示，但对它在DNA复制过程中的生化行为的理解仍然是一个谜。在这里，我们分析了染色质结合动力学的FACT使用非洲爪蟾卵提取物无细胞系统。我们发现FACT至少有两个不同的染色质结合阶段：（1）在DNA复制开始时的快速染色质结合阶段，不涉及起源许可;（2）在起源许可后开始的染色质结合的第二阶段。有趣的是，早期结合的FACT从染色质中解离时，DNA复制被阻断的Cdc 6在原始环之前的许可状态。Cdc 6诱导的去除FACT被阻断的抑制与双生子的起源许可，但不是通过抑制DNA聚合酶，CDK，或Cdc 7的活性。此外，染色质转移实验表明，损害FACT的后期结合严重损害DNA复制活性。两者合计，我们建议，即使FACT具有快速的染色质结合活性，FACT对染色质的结合模式的变化后，原产地许可，这可能有助于建立其功能链接的DNA复制机制。
The facilitates chromatin transcription (FACT) complex affects nuclear DNA transactions in a chromatin context. Though the involvement of FACT in eukaryotic DNA replication has been revealed, a clear understanding of its biochemical behavior during DNA replication still remains elusive. Here, we analyzed the chromatin-binding dynamics of FACT using Xenopus egg extract cell-free system. We found that FACT has at least two distinct chromatin-binding phases: (1) a rapid chromatin-binding phase at the onset of DNA replication that did not involve origin licensing and (2) a second phase of chromatin binding that initiated after origin licensing. Intriguingly, early-binding FACT dissociated from chromatin when DNA replication was blocked by the addition of Cdc6 in the licensed state before origin ring. Cdc6-induced removal of FACT was blocked by the inhibition of origin licensing with geminin, but not by suppressing the activity of DNA polymerases, CDK, or Cdc7. Furthermore, chromatin transfer experiments revealed that impairing the later binding of FACT severely compromises DNA replication activity. Taken together, we propose that even though FACT has rapid chromatin-binding activity, the binding pattern of FACT on chromatin changes after origin licensing, which may contribute to the establishment of its functional link to the DNA replication machinery.
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