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DNA中真实的孢子光产物损伤结构提示了一种识别基础。

The structure of an authentic spore photoproduct lesion in DNA suggests a basis for recognition.

作者信息

Singh Isha, Jian Yajun, Li Lei, Georgiadis Millie M

机构信息

Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.

出版信息

Acta Crystallogr D Biol Crystallogr. 2014 Mar;70(Pt 3):752-9. doi: 10.1107/S1399004713032987. Epub 2014 Feb 15.

Abstract

The spore photoproduct lesion (SP; 5-thymine-5,6-dihydrothymine) is the dominant photoproduct found in UV-irradiated spores of some bacteria such as Bacillus subtilis. Upon spore germination, this lesion is repaired in a light-independent manner by a specific repair enzyme: the spore photoproduct lyase (SP lyase). In this work, a host-guest approach in which the N-terminal fragment of Moloney murine leukemia virus reverse transcriptase (MMLV RT) serves as the host and DNA as the guest was used to determine the crystal structures of complexes including 16 bp oligonucleotides with and without the SP lesion at 2.14 and 1.72 Å resolution, respectively. In contrast to other types of thymine-thymine lesions, the SP lesion retains normal Watson-Crick hydrogen bonding to the adenine bases of the complementary strand, with shorter hydrogen bonds than found in the structure of the undamaged DNA. However, the lesion induces structural changes in the local conformation of what is otherwise B-form DNA. The region surrounding the lesion differs significantly in helical form from B-DNA, and the minor groove is widened by almost 3 Å compared with that of the undamaged DNA. Thus, these unusual structural features associated with SP lesions may provide a basis for recognition by the SP lyase.

摘要

孢子光产物损伤(SP;5-胸腺嘧啶-5,6-二氢胸腺嘧啶)是在紫外线照射的某些细菌(如枯草芽孢杆菌)孢子中发现的主要光产物。在孢子萌发时,这种损伤由一种特定的修复酶——孢子光产物裂解酶(SP裂解酶)以不依赖光的方式进行修复。在这项工作中,采用了一种主客体方法,其中莫洛尼鼠白血病病毒逆转录酶(MMLV RT)的N端片段作为主体,DNA作为客体,分别以2.14 Å和1.72 Å的分辨率确定了包含有和没有SP损伤的16 bp寡核苷酸的复合物的晶体结构。与其他类型的胸腺嘧啶-胸腺嘧啶损伤不同,SP损伤与互补链的腺嘌呤碱基保持正常的沃森-克里克氢键,但其氢键比未损伤DNA结构中的氢键短。然而,这种损伤会在原本为B型DNA的局部构象中诱导结构变化。损伤周围区域的螺旋形式与B-DNA有显著差异,与未损伤DNA相比,小沟加宽了近3 Å。因此,这些与SP损伤相关的不寻常结构特征可能为SP裂解酶的识别提供基础。

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