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首页> 外文期刊>Molecular and Cellular Biology >Mapping of a replication origin within the promoter region of two unlinked, abundantly transcribed actin genes of Physarum polycephalum.
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Mapping of a replication origin within the promoter region of two unlinked, abundantly transcribed actin genes of Physarum polycephalum.

机译：复制起点在两个未连接，大量转录的cephal头肌动蛋白基因的启动子区域内的定位。

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摘要

We analyzed the replication of two unlinked actin genes, ardB and ardC , which are abundantly transcribed in the naturally synchronous plasmodium of the slime mold Physarum polycephalum. Detection and size measurements of single-stranded nascent replication intermediates (RIs) demonstrate that these two genes are concomitantly replicated at the onset of the 3-h S phase and tightly linked to replication origins. Appearance of RIs on neutral-neutral two-dimensional gels at specific time points in early S phase and analysis of their structure confirmed these results and further established that, in both cases, an efficient, site-specific, bidirectional origin of replication is localized within the promoter region of the gene. We also determined similar elongation rates for the divergent replication forks of the ardC gene replicon. Finally, taking advantage of a restriction fragment length polymorphism, we studied allelic replicons and demonstrate similar localizations and a simultaneous firing of allelic replication origins. Computer search revealed a low level of homology between the promoters of ardB and ardC and, most notably, the absence of DNA sequences similar to the yeast autonomously replicating sequence consensus sequence in these Physarum origin regions. Our results with the ardB and ardC actin genes support the model of early replicating origins located within the promoter regions of abundantly transcribed genes in P. polycephalum.
机译：我们分析了两个未链接的肌动蛋白基因ardB和ardC的复制，它们在粘液霉菌多头cephal的自然同步疟原虫中大量转录。单链新生复制中间体（RI）的检测和大小测量表明，这两个基因在3-h S期开始时同时复制，并与复制起点紧密相连。 RI在S早期特定时间点在中性-中性二维凝胶上的出现及其结构分析证实了这些结果，并进一步证实，在两种情况下，有效的，特定于位点的双向复制起点均位于基因的启动子区域。我们还为ardC基因复制子的不同复制叉确定了相似的延伸率。最后，利用限制性片段长度多态性，我们研究了等位基因复制子，并证明了相似的定位和等位基因复制起点的同时触发。计算机搜索显示在这些Physarum起源区域中，ardB和ardC的启动子之间的同源性较低，最显着的是，没有类似于酵母自主复制序列共有序列的DNA序列。我们用ardB和ardC肌动蛋白基因进行的研究结果支持了早期复制起点的模型，该起点位于多头体育中转录丰富的基因的启动子区域内。

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