Crystallization and preliminary X-ray characterization of the eukaryotic replication terminator Reb1-Ter DNA complex.
Crystallization and preliminary X-ray characterization of the eukaryotic replication terminator Reb1-Ter DNA complex.
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DOI：
10.1107/s2053230x15004112
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发表时间：
2015-04
期刊：
影响因子：
--
通讯作者：
Escalante CR
中科院分区：
文献类型：
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作者：
Jaiswal R;Singh SK;Bastia D;Escalante CR
来自粟酒裂殖酵母的 Reb1 蛋白是控制真核细胞中程序性复制终止和/或转录终止的蛋白家族的成员。这些事件发生在天然存在的复制叉屏障 (RFB) 处，其中 Reb1 与终止 (Ter) DNA 位点结合，并协调复制叉的极性停滞和向相反方向接近的转录。 Reb1 DNA 结合和复制终止结构域在大肠杆菌中表达、纯化并与 26 聚体 DNA Ter 位点复合结晶。需要在油下批量结晶以产生用于数据收集的良好质量的晶体。晶体在 P21 空间群中生长，晶胞参数 a = 68.9、b = 162.9、c = 71.1 Å、β = 94.7°。晶体衍射分辨率为 3.0 Å。晶体是镶嵌的，需要两到三个退火周期。这项研究首次获得了有关这一重要蛋白质家族的结构信息，并将提供对复制和转录终止机制的见解。
The Reb1 protein from Schizosaccharomyces pombe is a member of a family of proteins that control programmed replication termination and/or transcription termination in eukaryotic cells. These events occur at naturally occurring replication fork barriers (RFBs), where Reb1 binds to termination (Ter) DNA sites and coordinates the polar arrest of replication forks and transcription approaching in opposite directions. The Reb1 DNA-binding and replication-termination domain was expressed in Escherichia coli, purified and crystallized in complex with a 26-mer DNA Ter site. Batch crystallization under oil was required to produce crystals of good quality for data collection. Crystals grew in space group P21, with unit-cell parameters a = 68.9, b = 162.9, c = 71.1 Å, β = 94.7°. The crystals diffracted to a resolution of 3.0 Å. The crystals were mosaic and required two or three cycles of annealing. This study is the first to yield structural information about this important family of proteins and will provide insights into the mechanism of replication and transcription termination.