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首页> 外文期刊>Journal of cellular biochemistry. >Major DNA replication initiation sites in the c-myc locus in human cells.
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Major DNA replication initiation sites in the c-myc locus in human cells.

机译：人细胞中c-myc基因座中的主要DNA复制起始位点。

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摘要

DNA replication initiation sites and initiation frequencies over 12. 5 kb of the human c-myc locus, including 4.6 kb of new 5' sequence, were determined based on short nascent DNA abundance measured by competitive polymerase chain reaction using 21 primer sets. In previous measurements, no comparative quantitation of nascent strand abundance was performed, and distinction of major from minor initiation sites was not feasible. Two major initiation sites were identified in this study. One predominant site has been located at approximately 0.5 kb upstream of exon 1 of the c-myc gene, and a second new major site is located in exon 2. The site in exon 2 has not been previously identified. In addition, there are other sites that may act as less frequently used initiation sites, some of which may correspond to sites in previous reports. Furthermore, a comparison of the abundance of DNA replication intermediates over this same region of the c-myc locus between HeLa and normal skin fibroblast (NSF) cells indicated that the relative distribution was very similar, but that nascent strand abundance in HeLa cells was approximately twice that in NSF relative to the abundance at the lamin B2 origin. This increased activity at initiation sites in the c-myc locus may mainly be influenced by regulators at higher levels in transformed cells like HeLa. Copyright 2000 Wiley-Liss, Inc.
机译：基于通过使用21个引物组的竞争性聚合酶链反应测量的短时新生DNA丰度，确定了超过12个人c-myc基因座的DNA复制起始位点和起始频率，包括4.6 kb的新5'序列。在以前的测量中，未对新生链的丰度进行比较定量，并且将主要和次要起始位点区分开是不可行的。这项研究确定了两个主要的起始位点。一个主要位点位于c-myc基因外显子1上游约0.5 kb，另一个新的主要位点位于外显子2。先前未鉴定外显子2中的位点。此外，还有其他站点可以充当不那么频繁使用的启动站点，其中一些站点可能对应于先前报告中的站点。此外，在HeLa细胞与正常皮肤成纤维细胞（NSF）细胞之间的c-myc基因座同一区域上的DNA复制中间体的丰度比较表明，相对分布非常相似，但HeLa细胞中新生链的丰度约为相对于Lamin B2来源的丰度，是NSF中的两倍。在c-myc基因座起始位点的这种增加的活性可能主要受转化细胞（如HeLa）中较高水平的调节剂的影响。版权所有2000 Wiley-Liss，Inc.

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