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首页> 外文期刊>Nucleic Acids Research >H-1 NMR STUDIES OF THE 5-(HYDROXYMETHYL)-2'-DEOXYURIDINE CONTAINING TF1 BINDING SITE
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H-1 NMR STUDIES OF THE 5-(HYDROXYMETHYL)-2'-DEOXYURIDINE CONTAINING TF1 BINDING SITE

机译：含TF1结合位点的5-（羟甲基）-2'-脱氧尿嘧啶的H-1 NMR研究

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摘要

The pyrimidine base 5-(hydroxymethyl)-2'-deoxyuridine (HmU) is a common nucleotide in SPO1 phage DNA. Numerous transcriptional proteins bind HmU-containing DNA preferentially implicating a regulatory function of HmU. We Rave investigated the conformation and dynamics of d-(5'-CHmUCHmUACACGHmUGHmUA-GAG-OH-3')(2) (HmU-DNA). This oligonucleotide mimics the consensus sequence of Transcription Factor 1 (TF1). The HmU-DNA was compared to the thymine-containing oligonucleotide. NOESY and DQF COSY spectroscopy provided resonance assignments of nonexchangeable and exchangeable protons, intranucleotide, internucleotide acid intrastrand proton-proton distances, and dihedral angle constraints. Methylene protons of the hydroxymethyl group are nonequivalent protons and the hydroxymethyl group is not freely rotating. The hydroxymethyl group adopts a specific orientation with the OH group oriented on the 3' side of the plane of the base, Analysis of imino proton resonances and NOEs indicates additional end base pair fraying and a temperature-induced transition to a conformation in which the internal HmU-A base pairs are disrupted or have reduced lifetimes. Orientation of the hydroxymethyl group indicates the presence of internucleotide intrastrand hydrogen bonding between the HmU12C5 hydroxyl group and A13, All sugars in both DNAs show a C2'endo conformation (typical of B-DNA).
机译：嘧啶碱基的5-（羟甲基）-2'-脱氧尿苷（HmU）是SPO1噬菌体DNA中的常见核苷酸。许多转录蛋白结合含HmU的DNA，优先牵涉HmU的调节功能。我们Rave研究了d-（5'-CHmUCHmUACACGHmUGHmUA-GAG-OH-3'）（2）（HmU-DNA）的构象和动力学。该寡核苷酸模拟转录因子1（TF1）的共有序列。将HmU-DNA与含胸腺嘧啶的寡核苷酸进行比较。 NOESY和DQF COZY光谱学提供了不可交换和可交换质子，核苷酸内，核苷酸间酸内链内质子-质子距离和二面角约束的共振分配。羟甲基的亚甲基质子为非等价的质子，羟甲基不自由旋转。羟甲基基团采用特定的取向，OH基团位于碱基平面的3'侧。对亚氨基质子共振和NOE的分析表明，额外的末端碱基对磨损以及温度诱导的构象转变，其中内部HmU-A碱基对被破坏或寿命缩短。羟甲基的方向指示在HmU12C5羟基和A13之间存在核苷酸间链内氢键。两个DNA中的所有糖均显示C2'内构象（典型为B-DNA）。

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