Site-specific oligodeoxynucleotide binding to maize Adh1 gene promoter represses Adh1-GUS gene expression in vivo.
Site-specific oligodeoxynucleotide binding to maize Adh1 gene promoter represses Adh1-GUS gene expression in vivo.
复制标题
位点特异性寡脱氧核苷酸与玉米 Adh1 基因启动子结合可抑制体内 Adh1-GUS 基因表达。
DOI：
10.1007/bf00027068
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发表时间：
1992
影响因子：
5.1
通讯作者：
Ferl,RJ
中科院分区：
文献类型：
--
作者：
Lu,G;Ferl,RJ
在maizeadh1基因启动子中存在一条36bp的极端同嘌呤/同嘧啶（PuPy）不对称链（从- 44到- 79），在超线圈张力下质粒对s1超敏感。设计了PuPy通道对应的寡脱氧核苷酸，以检查该区域的二级结构，并解决该通道在基因调控中的可能作用。通过对低聚脱氧核苷酸的带移和dna酶I足迹分析，得出了同嘧啶低聚脱氧核苷酸能与体外双链PuPy牵引力蛋白形成三螺旋结构的结论。原生质体、悬浮细胞和幼苗根的瞬时实验表明，同嘧啶寡脱氧核苷酸也能够在共转化过程中抑制adh1 - gus基因的表达，可能是通过与PuPy牵拉蛋白在体内形成三螺旋结构来实现的。互补的同嘌呤寡脱氧核苷酸在体外不会形成三重螺旋蛋白，在体内也不会抑制sadh1 - gusin。我们提出三螺旋形成是一种潜在的体内调节现象，并且区域内三螺旋可能通过H-DNA的形成在adh1启动子内发生。
There is a 36 bp tract of extreme homopurine/homopyrimidine (PuPy) asymmetry in the maizeAdh1gene promoter (from −44 to −79) that is S1-hypersensitive in plasmids under supercoil tension. Oligodeoxynucleotides corresponding to the PuPy tract were designed to examine the secondary structure of the region and address the possible role of the tract in gene regulation. On the basis of oligodeoxynucleotide band-shift and DNase I footprinting analyses, it was concluded that the homopyrimidine oligodeoxynucleotide can form a triple helix with the duplex PuPy tractin vitro. Transient assays in protoplasts, suspension cells, and seedling roots show that the homopyrimidine oligodeoxynucleotide is also capable of repressingAdh1-GUS gene expression during co-transformation, presumably by the formation of a triple helix with the PuPy tractin vivo. The complementary homopurine oligodeoxynucleotide would not form a triple helixin vitro, nor would it repressAdh1-GUSin vivo. We propose that triple helix formation is a potential regulatory phenomenonin vivo, and that an intraregion triple helix could occur within theAdh1promoter via the formation of H-DNA.