Efficient Synthesis of DNA Duplexes Containing Reduced Acetaldehyde Interstrand Cross-Links.
Efficient Synthesis of DNA Duplexes Containing Reduced Acetaldehyde Interstrand Cross-Links.
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DOI：
10.1021/jacs.2c10070
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发表时间：
2023-01-18
影响因子：
15
通讯作者：
Grasby, Jane A.
中科院分区：
文献类型：
--
作者：
Williams, David M.;Morton, Sally B.;Finger, L. David;van der Sluijs, Roxanne;Mulcrone, William D.;Hodskinson, Michael;Millington, Christopher L.;Vanhinsbergh, Christina;Patel, Ketan J.;Dickman, Mark J.;Knipscheer, Puck;Grasby, Jane A.
DNA链间交联（ICL）阻止DNA复制和转录，并可能导致潜在的致命事件，如癌症或骨髓衰竭。ICL通常由范可尼贫血（FA）通路内的蛋白质修复，尽管该通路的细节尚未完全确定。产生含有ICL的DNA的方法是进一步理解DNA交联修复的关键。体内ICL形成的主要途径包括DNA与乙醇代谢产生的乙醛反应。该反应形成三碳桥接的ICL，其涉及双链体的相对链中的相邻鸟嘌呤的氨基，从而产生氨基和亚氨基官能团。一个稳定的还原形式的ICL在理解这些类型的加合物的识别和修复中有应用。先前产生含有这些加合物的DNA双链体的途径涉及冗长的DNA合成后化学，随后还原亚胺。在这里，一个有效的和高产率的方法来减少ICL使用新的N2-（（R）-4-三氟乙酰氨基丁烷-2-基）-2′-脱氧鸟苷亚磷酰胺的描述。在标准的自动化DNA合成和脱保护之后，在室温下与含有2-氟-2 ′-脱氧肌苷的互补寡脱氧核糖核苷酸孵育过夜后，以超过90%的产率形成ICL。交联的双链体显示出比对照序列高25 °C的解链转变。重要的是，我们表明使用爪蟾卵提取物系统，通过这种方法合成的ICL修复的FA途径。这种方法的简单性和效率，用于制备减少乙醛ICLs将有助于访问这些DNA结构的未来研究的交联修复。
DNA interstrand cross-links (ICLs) prevent DNA replication and transcription and can lead to potentially lethal events, such as cancer or bone marrow failure. ICLs are typically repaired by proteins within the Fanconi Anemia (FA) pathway, although the details of the pathway are not fully established. Methods to generate DNA containing ICLs are key to furthering the understanding of DNA cross-link repair. A major route to ICL formation in vivo involves reaction of DNA with acetaldehyde, derived from ethanol metabolism. This reaction forms a three-carbon bridged ICL involving the amino groups of adjacent guanines in opposite strands of a duplex resulting in amino and imino functionalities. A stable reduced form of the ICL has applications in understanding the recognition and repair of these types of adducts. Previous routes to creating DNA duplexes containing these adducts have involved lengthy post-DNA synthesis chemistry followed by reduction of the imine. Here, an efficient and high-yielding approach to the reduced ICL using a novel N2-((R)-4-trifluoroacetamidobutan-2-yl)-2′-deoxyguanosine phosphoramidite is described. Following standard automated DNA synthesis and deprotection, the ICL is formed overnight in over 90% yield upon incubation at room temperature with a complementary oligodeoxyribonucleotide containing 2-fluoro-2′-deoxyinosine. The cross-linked duplex displayed a melting transition 25 °C higher than control sequences. Importantly, we show using the Xenopus egg extract system that an ICL synthesized by this method is repaired by the FA pathway. The simplicity and efficiency of this methodology for preparing reduced acetaldehyde ICLs will facilitate access to these DNA architectures for future studies on cross-link repair.
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DOI：
10.1126/science.1182372
发表时间：
2009-12-18
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Knipscheer P;Räschle M;Smogorzewska A;Enoiu M;Ho TV;Schärer OD;Elledge SJ;Walter JC
通讯作者：
Walter JC
DOI：
10.1038/nrc3088
发表时间：
2011-06-24
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
通讯作者：
--
影响因子：
4.1
作者：
Kozekov, ID;Nechev, LV;Harris, TM
通讯作者：
Harris, TM
影响因子：
15
作者：
Kozekov, ID;Nechev, LV;Harris, TM
通讯作者：
Harris, TM
影响因子：
4.1
作者：
Nechev, LV;Kozekov, I;Harris, TM
通讯作者：
Harris, TM