Effect of sequence context and direction of replication on AP site bypass in Saccharomyces cerevisiae.
Effect of sequence context and direction of replication on AP site bypass in Saccharomyces cerevisiae.
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DOI：
10.1016/j.mrfmmm.2009.06.006
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发表时间：
2009-10-02
影响因子：
2.3
通讯作者：
Kow, Yoke W.
中科院分区：
文献类型：
--
作者：
Bao, Gaobin;Kow, Yoke W.
关键词：
酵母可以很容易地被单链寡核苷酸（ssOligos）转化。之前，我们证明了生成包含 AP 位点的 1-nt 环的 ssOligo 纠正了 lys2ΔA746 等位基因中的 -1 移码突变。然而，这些实验必须在缺乏主要 AP 核酸内切酶的酵母 apn1 突变体中进行。在本研究中，我们表明可以通过使用 ssOligos 在修复能力强的酵母中研究 AP 位点的旁路，该 ssOligos 生成包含 AP 位点的 7-nt 环。使用生成 7-nt 环的 ssOligos 进行的旁路研究得到了验证，证明所获得的结果与使用 apn1 突变体中包含 1-nt 环的 ssOligos 得出的结果相似。通过使用 7-nt 环系统，我们表明 AP 位点的旁路效率取决于病变周围的序列背景，并且显然不受 DNA 复制方向的影响。相反，AP 位点的诱变特异性不受序列背景或复制方向的影响。在所有情况下，dC 的插入频率是 dA 与 AP 位点相对的频率的两倍，表明 REV1 主要负责在所有研究的病变位点绕过 AP 位点。
Yeast can be readily transformed by single-stranded oligonucleotides (ssOligos). Previously, we showed that an ssOligo that generates a 1-nt loop containing an AP site corrected the −1 frameshift mutation in the lys2ΔA746 allele. However, these experiments had to be performed in yeast apn1 mutants lacking the major AP endonuclease. In this study, we show that bypass of an AP site can be studied in repair-proficient yeast by using ssOligos that generates a 7-nt loop containing an AP site. The bypass studies performed using the ssOligos that generate a 7-nt loop was validated by demonstrating that the result obtained are similar to those derived using ssOligos containing a 1-nt loop in an apn1 mutant. By using the 7-nt loop system, we showed that the bypass efficiencies of AP sites are dependent on the sequence context that surrounds the lesion and are apparently no affected by the direction of DNA replication. In contrast, the mutagenic specificity of an AP site is not affected by the sequence context or the direction of replication. In all cases, dC is inserted at twice the frequency of dA opposite an AP site, indicating that REV1 is mainly responsible for bypass of AP sites at all lesion sites studied.
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影响因子：
2.9
作者：
Mekhovich, O;Tang, MS;Romano, LJ
通讯作者：
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影响因子：
5.6
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DOI：
10.1098/rstb.2000.0001
发表时间：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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