Central base pair flipping and discrimination by PspGI.

Central base pair flipping and discrimination by PspGI.
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DOI:
10.1093/nar/gkn622
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发表时间:
2008-11
影响因子:
14.9
通讯作者:
Bochtler, Matthias
Bochtler, Matthias
中科院分区:
生物学2区
文献类型:
--
作者:
Szczepanowski, Roman H.;Carpenter, Michael A.;Czapinska, Honorata;Zaremba, Mindaugas;Tamulaitis, Gintautas;Siksnys, Virginijus;Bhagwat, Ashok S.;Bochtler, Matthias

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PspGI是识别与CCNGG相关的五聚体序列的一组限制性内切核酸酶的代表。与先前研究的Ecl 18 kI不同,其对中心碱基对没有任何特异性,PspGI在其靶位点中偏好A/T而不是G/C。在这里,我们提出了一个结构的PspGI与目标DNA在1.7微米分辨率。在这种结构中,识别序列中心的碱基从DNA中挤出并翻转到PspGI的口袋中。翻转的胸腺嘧啶是通常的反构象,但翻转的腺嘌呤采取通常不利的顺式构象。这一结果和随附的手稿将翻转位置中A/T对优于G/C对的偏好归因于翻转A/T对的固有较低罚分以及针对鸟嘌呤和胞嘧啶的PspGI口袋的选择。我们的数据表明,翻转可以有助于正常碱基之间的歧视。这为碱基翻转增加了一个新的角色,除了它在碱基修饰和DNA损伤修复中的众所周知的功能。
PspGI is a representative of a group of restriction endonucleases that recognize a pentameric sequence related to CCNGG. Unlike the previously investigated Ecl18kI, which does not have any specificity for the central base pair, PspGI prefers A/T over G/C in its target site. Here, we present a structure of PspGI with target DNA at 1.7 Å resolution. In this structure, the bases at the center of the recognition sequence are extruded from the DNA and flipped into pockets of PspGI. The flipped thymine is in the usual anti conformation, but the flipped adenine takes the normally unfavorable syn conformation. The results of this and the accompanying manuscript attribute the preference for A/T pairs over G/C pairs in the flipping position to the intrinsically lower penalty for flipping A/T pairs and to selection of the PspGI pockets against guanine and cytosine. Our data show that flipping can contribute to the discrimination between normal bases. This adds a new role to base flipping in addition to its well-known function in base modification and DNA damage repair.
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