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Chlamydomonas reinhardtii telomere repeats form unstable structures involving guanine-guanine base pairs

机译:莱茵衣藻端粒重复序列形成不稳定结构,涉及鸟嘌呤-鸟嘌呤碱基对

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摘要

Unusual DNA structures involving four guanines in a planar formation (guanine tetrads) are formed by guanine-rich (G-rich) telomere DNA and other G-rich sequences (reviewed in (1)) and may be important in the structure and function of telomeres. These structures result from intrastrand and/or interstand Hoogsteen base pairs between the guanines. We used the telomeric repeat of Chlamydomonas reinhardtii, TTTTAGGG, which contains 3 guanines and has a long interguanine A + T tract, to determine whether these sequences can form intrastrand and interstrand guanine tetrads. We have found that ss (TTTTAGGG)4 can form intrastrand guanine tetrads that are less stable than those formed by more G-rich telomere sequences. They are not only more stable, but also more compact, they are more stable in the presence of K+ than they are in the presence of Na+. While ds oligonucleotides with ss 3' overhangs of (TTTTAGGG)2 can be observed to associate as dimers, formation of this interstrand guanine tetrad structure occurs to a very limited extent and requires very high G-strand concentration, high ionic strength, and at least 49 hours of incubation. Our results suggest that, if telomere dimerization occurs in vivo, it would require factors in addition to the TTTTAGGG telomere sequence.
机译:富含鸟嘌呤(G富集)端粒DNA和其他富含G的序列形成了不寻常的涉及四个鸟嘌呤的平面结构(鸟嘌呤四联体)(在(1)中进行了综述),并且可能对鸟嘌呤的结构和功能很重要端粒。这些结构由鸟嘌呤之间的链内和/或链节间Hoogsteen碱基对产生。我们使用了莱茵衣藻TTTTAGGG的端粒重复序列,该序列包含3个鸟嘌呤并且具有较长的鸟嘌呤间A + T链,来确定这些序列是否可以形成链内和链间鸟嘌呤四联体。我们发现,ss(TTTTAGGG) 4 可以形成链内鸟嘌呤四联体,其稳定性不如由富含G的端粒序列形成的那些。它们不仅更稳定,而且更紧凑,在K + 存在下比在Na + 存在下更稳定。尽管可以观察到具有(TTTTAGGG) 2 的ss 3'突出端的ds寡核苷酸以二聚体缔合,但这种链间鸟嘌呤四联体结构的形成非常有限,并且需要非常高的G链浓度,高离子强度,并且至少需要49小时的孵育时间。我们的结果表明,如果端粒在体内发生二聚化,则除了TTTTAGGG端粒序列外,还需要其他因素。

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