Formation of the G-quadruplex and i-motif structures in retinoblastoma susceptibility genes (Rb).

Formation of the G-quadruplex and i-motif structures in retinoblastoma susceptibility genes (Rb).
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DOI:
10.1093/nar/gkj485
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发表时间:
2006
影响因子:
14.9
通讯作者:
Sugiyama H
Sugiyama H
中科院分区:
生物学2区
文献类型:
--
作者:
Xu Y;Sugiyama H

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用化学修饰、圆二色性(CD)和荧光光谱法研究了视网膜母细胞瘤(Rb)基因5′端G-四链体和i-基序结构的形成。结果发现,在反平行G-四链体中显示顺式构象的位置处的8-甲基鸟嘌呤的取代稳定了富含G链的结构。互补的富含C的18聚体形成i基序结构,如CD光谱所示。基于C到T突变实验,确定了参与i-基序结构的C-C+碱基对的C碱基。2-氨基嘌呤(2-AP)取代实验表明,相对于双链体,G-四链体中荧光的增加归因于G-四链体环内未堆叠的2-AP。荧光实验表明,G-四链体和i-基序的形成可以与双链体形成竞争。此外,聚合酶阻滞试验表明,在Rb基因中形成的G-四链体结构作为DNA合成的障碍。
The formation of G-quadruplex and i-motif structures in the 5′ end of the retinoblastoma (Rb) gene was examined using chemical modifications, circular dichroism (CD) and fluorescence spectroscopy. It was found that substitutions of 8-methylguanine at positions that show syn conformations in antiparallel G-quadruplexes stabilize the structure in the G-rich strand. The complementary C-rich 18mer forms an i-motif structure, as suggested by CD spectroscopy. Based on the C to T mutation experiments, C bases participated in the C–C+ base pair of the i-motif structure were determined. Experiments of 2-aminopurine (2-AP) substitution reveal that an increase of fluorescence in the G-quadruplex relative to duplex is attributed to unstacked 2-AP within the loop of G-quadruplex. The fluorescence experiments suggest that formation of the G-quadruplex and i-motif can compete with duplex formation. Furthermore, a polymerase arrest assay indicated that formation the G-quadruplex structure in the Rb gene acts as a barrier in DNA synthesis.
DOI: 10.1038/nsb0294-83
发表时间: 1994-02-01
期刊: NATURE STRUCTURAL BIOLOGY
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