d'Ambrosio E, Furano A V
Nucleic Acids Res. 1987 Apr 10;15(7):3155-75. doi: 10.1093/nar/15.7.3155.
An approximately equal to 150-bp GC-rich (approximately equal to 60%) region is at the right end of rat long interspersed repeated DNA (LINE or L1Rn) family members. We report here that one of the DNA strands from this region contains several non-palindromic sites that strongly arrest DNA synthesis in vitro by the prokaryotic Klenow and T4 DNA polymerases, the eukaryotic alpha polymerase, and AMV reverse transcriptase. The strongest arrest sites are G-rich (approximately equal to 70%) homopurine stretches of 18 or more residues. Shorter homopurine stretches (12 residues or fewer) did not arrest DNA synthesis even if the stretch contains 11/12 G residues. Arrest of the prokaryotic polymerases was not affected by their respective single strand binding proteins or polymerase accessory proteins. The region of duplex DNA which contains DNA synthesis arrest sites reacts with bromoacetaldehyde when present in negatively supercoiled molecules. By contrast, homopurine stretches that do not arrest DNA synthesis do not react with bromoacetaldehyde. The presence of bromoacetaldehyde-reactive bases in a G-rich homopurine-containing duplex under torsional stress is thought to be caused by base stacking in the homopurine strand. Therefore, we suggest that base-stacked regions of the template arrest DNA synthesis.
在大鼠长散在重复DNA(LINE或L1Rn)家族成员的右端有一个约150bp富含GC(约60%)的区域。我们在此报告,该区域的一条DNA链含有几个非回文位点,这些位点在体外能强烈抑制原核生物的Klenow和T4 DNA聚合酶、真核生物的α聚合酶以及禽成髓细胞瘤病毒逆转录酶的DNA合成。最强的抑制位点是富含G(约70%)的18个或更多残基的同型嘌呤序列。较短的同型嘌呤序列(12个残基或更少)即使包含11/12个G残基也不会抑制DNA合成。原核生物聚合酶的抑制不受其各自的单链结合蛋白或聚合酶辅助蛋白的影响。当存在于负超螺旋分子中时,包含DNA合成抑制位点的双链DNA区域会与溴乙醛发生反应。相比之下,不抑制DNA合成的同型嘌呤序列不会与溴乙醛发生反应。富含G的含同型嘌呤双链在扭转应力下存在与溴乙醛反应的碱基,被认为是由同型嘌呤链中的碱基堆积引起的。因此,我们认为模板的碱基堆积区域会抑制DNA合成。