Effect of nuclear environment on the distribution of benzo[a]pyrene diol epoxide-induced adducts in the HPRT gene of human fibroblasts.
Effect of nuclear environment on the distribution of benzo[a]pyrene diol epoxide-induced adducts in the HPRT gene of human fibroblasts.
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核环境对苯并[a]芘二醇环氧化物诱导的加合物在人成纤维细胞HPRT基因中分布的影响。
DOI：
10.1093/carcin/17.12.2695
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发表时间：
1996
期刊：
影响因子：
4.7
通讯作者：
McCormick,JJ
中科院分区：
文献类型：
--
作者：
Wei,D;Maher,VM;McCormick,JJ
（±）- 7β， 8α -二羟基- 9α， 10α-环氧- 7,8,9,10 -四氢苯并[a]芘（BPDE）是致癌性环境污染物苯并[a]芘的主要反应代谢物。与无蛋白DNA相比，bpde诱导的加合物形成在染色质DNA中的分布已经进行了深入的研究。然而，直到最近，在完整的哺乳动物细胞中，在核苷酸水平上对bpde诱导的加合物形成的研究还不可行。我们使用连接介导的聚合酶链反应（LMPCR）结合大肠杆菌（escherichia coliUvrABC）切切酶，在同步细胞群中从单个核苷酸到单个核苷酸的水平上，研究了bpde诱导的加合物在正常人hprt基因外显子3非转录链上的分布。我们发现，在G1期早期、s期、G2/ m期晚期和中期阻断的细胞中，BPDE加合物在目标区域的相对分布基本上是相同的。此外，对于几乎所有核苷酸位置，完整细胞中BPDE加合物的相对分布与纯化DNA在体外用BPDE处理时发现的非常相似。唯一的例外是，在体内，与体外处理的DNA相比，六个连续鸟嘌呤（即核苷酸207-212）区域的加合物形成强烈增强。利用测序酶和UvrABC切酶对bpde诱导的加合物形成进行了体外研究，结果表明，这个由6个连续的鸟嘌呤组成的区域采用了一种特殊的DNA构象。因此，我们得出结论，bpde诱导的核苷酸207 - 212加合物形成的增强不是反映蛋白质-DNA相互作用，而是反映了细胞核内生理环境对局部DNA构象的影响，并且这种影响在整个细胞周期中保持不变。
(±) - 7β, 8α - Dihydroxy - 9α, 10α-epoxy - 7, 8, 9, 10 - tetrahydro-benzo[a]pyrene (BPDE) is the principal reactive metabolite of the carcinogenic environmental pollutant benzo[a]pyr-ene. Intensive studies of the distribution of BPDE-induced adduct formation in chromatin DNA compared to that in protein-free DNA have been conducted. However, until recently, investigation of BPDE-induced adduct formation at the nucleotide level in intact mammalian cells has not been feasible. We used ligation-mediated polymerase chain reaction (LMPCR) in conjunction withEscherichia coliUvrABC excinuclease to investigate the distribution of BPDE-induced adducts in the non-transcribed strand of exon 3 of theHPRTgene in normal human fib rob lasts at the level of individual nucleotides to single nucleotide resolution using synchronized cell populations. We found that the relative distribution of BPDE adducts in the region of interest was essentially the same in cells treated in early G1 phase, S-phase, late G2/Mphase, and in cells blocked at metaphase. Furthermore, for almost all nucleotide positions, the relative distribution of BPDE adducts in the intact cells was very similar to that found when purified DNA was treated with BPDEin vitro. The only exception was thatin vivo, adduct formation at a region of six consecutive guanines, i.e. nucleotides 207–212, was strongly enhanced compared with that seen with DNA treatedin vitro. No obvious nucleosomal structures or other protein-DNA interaction were detected within the region of interest byin vivofoot printing with micrococcal nuclease and other reagents revealedIn vitrostudies mapping BPDE-induced adduct formation using Sequenase and UvrABC excinuclease suggested that this region of six consecutive guanines adopts a special DNA conformation. Therefore, we conclude that rather than reflecting protein-DNA interaction, the enhanced BPDE-induced adduct formation at nucleotides 207–212in vivoreflects the impact of the physiological environment in the cell nucleus on the local DNA conformation, and that this effect remains constant throughout the cell cycle.
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DOI：
10.1073/pnas.92.6.2204
发表时间：
1995-03
影响因子：
11.1
作者：
D. Wei;V. Maher;J. Mccormick
通讯作者：
D. Wei;V. Maher;J. Mccormick
致癌物质苯并[a]芘二醇环氧化物与重构为核小体的非洲爪蟾 5S DNA 共价结合。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Smith,BL;MacLeod,MC
通讯作者：
MacLeod,MC
影响因子：
56.9
作者：
PFEIFER, GP;STEIGERWALD, SD;RIGGS, AD
通讯作者：
RIGGS, AD
猿猴病毒 40 染色质超敏感区域附近定位核小体的体外组装。
影响因子：
5.6
作者：
J. H. Powers;M. Bina
通讯作者：
M. Bina
通过用 aphidicolin 抑制 DNA 聚合酶 α 来同步 HeLa 细胞培养物。
影响因子：
14.9
作者：
G. Pedrali;Silvio Spadari;A. Miller;A.O.A. Miller;J. Kruppa;G. Koch
通讯作者：
G. Koch