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位点特异性修饰的寡脱氧核糖核苷酸作为大肠杆菌DNA聚合酶I的模板。

Site-specifically modified oligodeoxyribonucleotides as templates for Escherichia coli DNA polymerase I.

作者信息

O'Connor D, Stöhrer G

出版信息

Proc Natl Acad Sci U S A. 1985 Apr;82(8):2325-9. doi: 10.1073/pnas.82.8.2325.

DOI:10.1073/pnas.82.8.2325
PMID:3887400
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC397550/
Abstract

Oligodeoxyribonucleotides with site-specific modifications have been used as substrates for Escherichia coli DNA polymerase I holoenzyme and Klenow fragment. Modifications included the bulky guanine-8-aminofluorene adduct and a guanine oxidation product resembling the product of photosensitized DNA oxidation. By a combination of primers and "nick-mers", conditions of single-strand-directed DNA synthesis and nick-translation could be created. Our results show that the polymerase can bypass both types of lesions. Bypass occurs on a single-stranded template but is facilitated on a nicked, double-stranded template. Only purines, with guanine more favored than adenine, are incorporated across both lesions. Hesitation during bypass could not be detected. The results indicate that site-specifically modified oligonucleotides can be sensitive probes for the action of polymerases on damaged templates. They also suggest a function for polymerase I, in its nick-translation capacity, during DNA repair and mutagenesis.

摘要

具有位点特异性修饰的寡脱氧核糖核苷酸已被用作大肠杆菌DNA聚合酶I全酶和克列诺片段的底物。修饰包括庞大的鸟嘌呤-8-氨基芴加合物和一种类似于光致敏DNA氧化产物的鸟嘌呤氧化产物。通过引物和“缺口寡核苷酸”的组合，可以创建单链定向DNA合成和缺口平移的条件。我们的结果表明，聚合酶可以绕过这两种类型的损伤。绕过发生在单链模板上，但在有缺口的双链模板上更容易进行。只有嘌呤可以跨过这两种损伤掺入，其中鸟嘌呤比腺嘌呤更受青睐。在绕过过程中未检测到犹豫现象。结果表明，位点特异性修饰的寡核苷酸可以作为聚合酶在受损模板上作用的敏感探针。它们还表明了聚合酶I在DNA修复和诱变过程中以其缺口平移能力所发挥的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/b690d49700ec/pnas00348-0139-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/f1e192ffe12c/pnas00348-0137-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/81f7f1980744/pnas00348-0137-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/73323c9c0599/pnas00348-0138-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/19971531149d/pnas00348-0138-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/b690d49700ec/pnas00348-0139-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/f1e192ffe12c/pnas00348-0137-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/81f7f1980744/pnas00348-0137-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/73323c9c0599/pnas00348-0138-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/19971531149d/pnas00348-0138-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/251a/397550/b690d49700ec/pnas00348-0139-a.jpg

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本文引用的文献

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Carcinogenesis. 1982;3(1):81-8. doi: 10.1093/carcin/3.1.81.
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Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli.大肠杆菌中紫外线和化学诱变所需基因产物的可诱导性。
Proc Natl Acad Sci U S A. 1981 Sep;78(9):5749-53. doi: 10.1073/pnas.78.9.5749.
3
Site-specific modification of the lactose operator with acetylaminofluorene.
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Proc Natl Acad Sci U S A. 1986 Jul;83(13):4599-603. doi: 10.1073/pnas.83.13.4599.
用乙酰氨基芴对乳糖操纵基因进行位点特异性修饰。
Nucleic Acids Res. 1983 Aug 11;11(15):5093-102. doi: 10.1093/nar/11.15.5093.
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Solid phase synthesis of polynucleotides. VIII. Synthesis of mixed oligodeoxyribonucleotides by the phosphotriester solid phase method.多核苷酸的固相合成。VIII. 用磷酸三酯固相法合成混合寡脱氧核糖核苷酸。
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