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Arabidopsis DNA Replication Initiates in Intergenic, AT-Rich Open Chromatin.
Plant Physiology ( IF 6.5 ) Pub Date : 2020-03-23 , DOI: 10.1104/pp.19.01520 Emily Wheeler 1 , Ashley M Brooks 2 , Lorenzo Concia 2 , Daniel L Vera 3 , Emily E Wear 2 , Chantal LeBlanc 4 , Umamaheswari Ramu 4 , Matthew W Vaughn 5 , Hank W Bass 6 , Robert A Martienssen 4 , William F Thompson 2 , Linda Hanley-Bowdoin 2
Plant Physiology ( IF 6.5 ) Pub Date : 2020-03-23 , DOI: 10.1104/pp.19.01520 Emily Wheeler 1 , Ashley M Brooks 2 , Lorenzo Concia 2 , Daniel L Vera 3 , Emily E Wear 2 , Chantal LeBlanc 4 , Umamaheswari Ramu 4 , Matthew W Vaughn 5 , Hank W Bass 6 , Robert A Martienssen 4 , William F Thompson 2 , Linda Hanley-Bowdoin 2
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The selection and firing of DNA replication origins play key roles in ensuring that eukaryotes accurately replicate their genomes. This process is not well documented in plants due in large measure to difficulties in working with plant systems. We developed a new functional assay to label and map very early replicating loci that must, by definition, include at least a subset of replication origins. Arabidopsis (Arabidopsis thaliana) cells were briefly labeled with 5-ethynyl-2'-deoxy-uridine, and nuclei were subjected to two-parameter flow sorting. We identified more than 5500 loci as initiation regions (IRs), the first regions to replicate in very early S phase. These were classified as strong or weak IRs based on the strength of their replication signals. Strong initiation regions were evenly spaced along chromosomal arms and depleted in centromeres, while weak initiation regions were enriched in centromeric regions. IRs are AT-rich sequences flanked by more GC-rich regions and located predominantly in intergenic regions. Nuclease sensitivity assays indicated that IRs are associated with accessible chromatin. Based on these observations, initiation of plant DNA replication shows some similarity to, but is also distinct from, initiation in other well-studied eukaryotic systems.
中文翻译：
拟南芥DNA复制始于富含AT的基因间开放染色质。
DNA复制起点的选择和激发在确保真核生物准确复制其基因组中起关键作用。由于在很大程度上难以操作工厂系统，因此该过程在工厂中没有得到很好的记录。我们开发了一种新的功能测定法来标记和定位非常早期的复制基因座，根据定义，该基因座必须至少包含复制起点的一个子集。用5-乙炔基-2'-脱氧尿苷短暂标记拟南芥（Arabidopsis thaliana）细胞，并对细胞核进行两参数流分选。我们确定了超过5500个基因座作为起始区域（IR），这是在非常早期的S期复制的第一个区域。根据其复制信号的强度，将它们分为强或弱红外。强启动区沿染色体臂均匀分布，并着落于着丝粒，而弱启动区则富集在着丝粒区。IR是富含AT的序列，其侧翼是更多的富含GC的区域，并且主要位于基因间区域。核酸酶敏感性测定表明IR与可及的染色质相关。基于这些观察，植物DNA复制的起始与其他经过充分研究的真核系统的起始相似，但也有区别。
更新日期：2020-05-01
中文翻译：
拟南芥DNA复制始于富含AT的基因间开放染色质。
DNA复制起点的选择和激发在确保真核生物准确复制其基因组中起关键作用。由于在很大程度上难以操作工厂系统，因此该过程在工厂中没有得到很好的记录。我们开发了一种新的功能测定法来标记和定位非常早期的复制基因座，根据定义，该基因座必须至少包含复制起点的一个子集。用5-乙炔基-2'-脱氧尿苷短暂标记拟南芥（Arabidopsis thaliana）细胞，并对细胞核进行两参数流分选。我们确定了超过5500个基因座作为起始区域（IR），这是在非常早期的S期复制的第一个区域。根据其复制信号的强度，将它们分为强或弱红外。强启动区沿染色体臂均匀分布，并着落于着丝粒，而弱启动区则富集在着丝粒区。IR是富含AT的序列，其侧翼是更多的富含GC的区域，并且主要位于基因间区域。核酸酶敏感性测定表明IR与可及的染色质相关。基于这些观察，植物DNA复制的起始与其他经过充分研究的真核系统的起始相似，但也有区别。
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