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首页> 外文期刊>Biochemistry >Cyclobutane thymine dimers with a disrupted phosphodiester bond are refractory to T4 endonuclease V digestion but have increased sensitivity to UvrABC nuclease
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Cyclobutane thymine dimers with a disrupted phosphodiester bond are refractory to T4 endonuclease V digestion but have increased sensitivity to UvrABC nuclease

机译：磷酸二酯键被破坏的环丁烷胸腺嘧啶二聚体对T4内切核酸酶V的消化是难治的，但对UvrABC核酸酶的敏感性增加

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UV irradiation induces the dimerization of synthetic single-stranded, 80-mer oligonucleotides with self-complementary, alternating purine-pyrimidine sequences, and terminal 5'- and 3'-thymines; this process can be reversed by photoreactivation. The UV-induced 160-mers are sensitive to digestion by the restriction enzyme SnaBI, but monomers are insensitive to digestion, indicating that UV irradiation stabilizes the formation of double-stranded DNA. These results suggest that UV irradiation of these 80-mer oligonucleotide substrates induces the formation of a novel cyclobutane thymine dimer which lacks an intradimer phosphodiester bond (CPD*). This CPD*, Linking the terminal thymines of two separate 80-mer molecules, is formed in a double-stranded DNA region created by self-annealing and intermolecular hybridization of the two 80-mer strands. We have found that these UV-induced CPD* in 160-mers are sensitive to cleavage by the nucleotide excision enzyme complex UvrABC nuclease, but resistant to cleavage by the cyclobutane pyrimidine dimer-specific enzyme T4 endonuclease V. However, pretreatment of the 160-mers with ligase reverses their sensitivity to these two enzymes, significantly reducing their susceptibility to cleavage by UvrABC nuclease but dramatically increasing their susceptibility to cleavage by T4 endonuclease. The biological significance of these findings is discussed. [References: 29]
机译：紫外线照射诱导具有自身互补的嘌呤-嘧啶序列以及末端5'-和3'-胸腺嘧啶的合成单链80-mer寡核苷酸的二聚化;此过程可以通过光活化逆转。紫外线诱导的160-mers对限制酶SnaBI的消化敏感，但是单体对消化不敏感，这表明紫外线辐射可以稳定双链DNA的形成。这些结果表明，这些80-聚体寡核苷酸底物的紫外线辐射诱导了新型二环丁烷胸腺嘧啶二聚体的形成，该环丁烷胸腺嘧啶二聚体缺乏二聚体内磷酸二酯键（CPD *）。连接两个单独的80-mer分子的末端胸腺嘧啶的CPD *在通过两条80-mer链的自退火和分子间杂交形成的双链DNA区域中形成。我们发现这些紫外线诱导的CPD *在160个单体中对核苷酸切除酶复合物UvrABC核酸酶的裂解敏感，但对环丁烷嘧啶二聚体特异性酶T4核酸内切酶V的裂解具有抵抗力。但是，对160-具有连接酶的聚合物逆转了它们对这两种酶的敏感性，显着降低了其对UvrABC核酸酶切割的敏感性，但显着提高了其对T4核酸内切酶切割的敏感性。讨论了这些发现的生物学意义。 [参考：29]

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