DEPURINATION-INDUCED INFIDELITY OF DEOXYRIBONUCLEIC-ACID SYNTHESIS WITH PURIFIED DEOXYRIBONUCLEIC-ACID REPLICATION PROTEINS INVITRO
DEPURINATION-INDUCED INFIDELITY OF DEOXYRIBONUCLEIC-ACID SYNTHESIS WITH PURIFIED DEOXYRIBONUCLEIC-ACID REPLICATION PROTEINS INVITRO
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DOI：
10.1021/bi00279a012
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发表时间：
1983-01-01
期刊：
影响因子：
2.9
通讯作者：
LOEB, LA
中科院分区：
文献类型：
--
作者：
KUNKEL, TA;SCHAAPER, RM;LOEB, LA
当用纯化的DNA聚合酶体外复制该DNA时，从v φ X174单链DNA中除去嘌呤碱基导致琥珀突变的回复频率增加。这种脱嘌呤诱导的诱变在3个不同的遗传基因座和几种不同的纯化酶中观察到，所述酶包括大肠杆菌DNA聚合酶I和III、禽成髓细胞瘤病毒DNA聚合酶和真核DNA聚合酶α，β。和γ。诱变的程度与病变旁路的估计频率相关，并且在缺乏校正能力的固有不准确的DNA聚合酶中最大。用大肠大肠杆菌DNA聚合酶I，条件下，减少校对的结果在3-5倍的增加脱嘌呤诱导的诱变，这表明了一个作用，校对在确定频率的旁路脱嘌呤位点。对E.大肠杆菌单链DNA结合蛋白对聚合酶I催化的反应与脱嘌呤DNA的诱变程度没有影响。在脱嘌呤的v φ X174琥珀3 DNA上，通过聚合酶I或禽成髓细胞瘤病毒DNA聚合酶在体外DNA合成期间产生的野生型回复突变体的分析表明，dAMP优选插入与推定的脱嘌呤位点587相对的位置。讨论了高等生物中嘌呤位点的致突变潜力和易错DNA合成的研究。
Removal of purine bases from .vphi.X174 single-stranded DNA leads to increased reversion frequency of amber mutations when this DNA is copied in vitro with purified DNA polymerases. This depurination-induced mutagenesis is observed at 3 different genetic loci and with several different purified enzymes, including Escherichia coli DNA polymerases I and III, avian myeloblastosis virus DNA polymerase and eukaryotic DNA polymerases .alpha., .beta. and .gamma.. The extent of mutagenesis correlates with the estimated frequency of bypass of the lesion and is greatest with inherently inaccurate DNA polymerases which lack proofreading capacity. With E. coli DNA polymerase I, conditions which diminish proofreading result in a 3-5-fold increase in depurination-induce mutagenesis, suggesting a role for proofreading in determining the frequency of bypass of apurinic sites. The addition of E. coli single-stranded DNA-binding protein to polymerase I catalyzed reactions with depurinated DNA had no effect of the extent of mutagenesis. Analysis of wild-type revertants produced during in vitro DNA synthesis by polymerase I or avian myeloblastosis virus DNA polymerase on depurinated .vphi.X174 amber 3 DNA indicates a preference for insertion of dAMP opposite the putative apurinic site at position 587. Both the mutagenic potential of a purinic sites in higher organisms and studies on error-prone DNA synthesis are discussed.