Solid phase synthesis and restriction endonuclease cleavage of oligodeoxynucleotides containing 5-(hydroxymethyl)-cytosine
Solid phase synthesis and restriction endonuclease cleavage of oligodeoxynucleotides containing 5-(hydroxymethyl)-cytosine
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DOI：
10.1093/nar/25.3.553
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发表时间：
1997-02-01
影响因子：
14.9
通讯作者：
Sowers, LC
中科院分区：
文献类型：
--
作者：
TardyPlanechaud, S;Fujimoto, J;Sowers, LC
新数据表明胞嘧啶甲基化在肿瘤发生中发挥重要作用。同时，最近的研究表明内源性 DNA 氧化损伤对人类疾病的发展起着重要作用。 DNA 中 5-甲基胞嘧啶 (5(m)C) 残基的 5-甲基氧化导致形成 5-(羟甲基)胞嘧啶 (C-hm)。脊椎动物 DNA 中 C-hm 残基的生物学后果尚不清楚。然而，疏水性甲基基团转化为亲水性羟甲基基团可能会显着改变序列特异性结合蛋白与DNA的相互作用。对于特定DNA损伤产物（例如C-hm）的潜在后果的生物物理和生物化学研究的核心是在选定位置合成含有此类修饰碱基的寡脱氧核苷酸的有效方法。在本文中，我们描述了一种使用已建立的亚磷酰胺化学将C-hm残基放置在寡脱氧核苷酸中的方法。此外，我们还研究了特定 C-hm 残基对甲基化敏感限制性内切酶 Msp1 和 Hpall 酶促切割寡脱氧核苷酸的影响。
Emerging data suggest an important role for cytosine methylation in tumorigenesis. Simultaneously, recent studies indicate a significant contribution of endogenous oxidative DNA damage to the development of human disease. Oxidation of the 5-methyl group of 5-methylcytosine (5(m)C) residues in DNA results in the formation of 5-(hydroxymethyl)cytosine (C-hm). The biological consequences of C-hm residues in vertebrate DNA are as yet unknown; however, conversion of the hydrophobic methyl group to the hydrophilic hydroxymethyl group may substantially alter the interaction of sequence-specific binding proteins with DNA, Central to both biophysical and biochemical studies on the potential consequences of specific DNA damage products such as C-hm are efficient methods for the synthesis of oligodeoxynucleotides containing such modified bases at selected positions, In this paper, we describe a method for the placement of C-hm residues in oligodeoxynucleotides using established phosphoramidite chemistry. In addition, we have examined the influence of specific C-hm residues on enzymatic cleavage of oligodeoxynucleotides by the methylation-sensitive restriction endonucleases Mspl and Hpall.