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首页> 外文期刊>Biochemistry >Miscoding properties of model estrogen-DNA adducts in reactions catalyzed by mammalian and Escherichia coli DNA polymerases.
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Miscoding properties of model estrogen-DNA adducts in reactions catalyzed by mammalian and Escherichia coli DNA polymerases.

机译：在哺乳动物和大肠杆菌DNA聚合酶催化的反应中，模型雌激素-DNA加合物的编码错误。

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摘要

The miscoding properties of the model estrogen-derived DNA adducts, N2-[3-methoxyestra-1,3,5(10)-trien-6-yl]-2'-deoxyguanosine (dG-N2-3MeE) and N6-[3-methoxyestra-1,3,5(10)-trien-6-yl]-2'- deoxyadenosine (dA-N6-3MeE), have been explored, using an in vitro experimental system to quantify base substitutions and deletions. Site-specifically modified oligodeoxynucleotides containing a single dG-N2-3MeE or dA-N6-3MeE were prepared postsynthetically and used as templates in primer extension reactions catalyzed by Escherichia coli and mammalian DNA polymerases. When the 3'-->5' exonuclease free (exo-) Klenow fragment of DNA polymerase I was used, dG-N2-3MeE promoted mostly one- and two-base deletions, along with small amounts of incorporation of dAMP, dGMP, and dCMP opposite the lesion. dA-N6-3MeE promoted the incorporation of dTMP opposite the lesion as well as two-base deletions, accompanied by the incorporation of dAMP. Using pol alpha, primer extension reactions were blocked at dG-N2-3MeE; however, dA-N6-3MeE promoted preferential incorporation of dTMP opposite the lesion with small amounts of incorporation of dCMP and deletions. Primer extension reactions catalyzed by pol delta were blocked at these lesions. When pol beta was used, dG-N2-3MeE produced small amounts of incorporation of dAMP and deletions. dA-N6-3MeE promoted preferential incorporation of dTMP, along with incorporation of dCMP and two-base deletions. The miscoding specificities and frequencies varied depending on the DNA polymerase used. These results indicate that estrogen-DNA adducts have miscoding potential.
机译：模型雌激素衍生的DNA加合物，N2- [3-甲氧基-1,3,5（10）-三烯-6-基] -2'-脱氧鸟苷（dG-N2-3MeE）和N6- [已使用体外实验系统对3-甲氧基es-1,3,5（10）-三烯-6-基] -2'-脱氧腺苷（dA-N6-3MeE）进行了定量，以定量碱基的取代和缺失。合成后制备了含有单个dG-N2-3MeE或dA-N6-3MeE的位点特异性修饰的寡脱氧核苷酸，并用作大肠杆菌和哺乳动物DNA聚合酶催化的引物延伸反应的模板。当使用DNA聚合酶I的无3'-> 5'核酸外切酶（exo-）Klenow片段时，dG-N2-3MeE主要促进一碱基和两碱基的缺失，以及少量dAMP，dGMP的掺入，病灶对面的dCMP。 dA-N6-3MeE促进了与病变相对的dTMP的掺入以及两个碱基的缺失，并伴有dAMP的掺入。使用pol alpha，引物延伸反应在dG-N2-3MeE处被阻断;然而，dA-N6-3MeE促进了病灶对面的dTMP的优先掺入以及少量的dCMP掺入和缺失。在这些病变处阻止了由polδ催化的引物延伸反应。使用pol beta时，dG-N2-3MeE产生了少量的dAMP掺入和缺失。 dA-N6-3MeE促进了dTMP的优先掺入，以及dCMP和两碱基缺失的掺入。误编码的特异性和频率取决于所用的DNA聚合酶。这些结果表明雌激素-DNA加合物具有错误编码的潜力。

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