دورية أكاديمية
In vivo mutational characterization of DndE involved in DNA phosphorothioate modification.
| العنوان: | In vivo mutational characterization of DndE involved in DNA phosphorothioate modification. |
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| المؤلفون: | Chongde Lai, Xiaolin Wu, Chao Chen, Teng Huang, Xiaolin Xiong, Shuangju Wu, Meijia Gu, Zixin Deng, Xi Chen, Shi Chen, Lianrong Wang |
| المصدر: | PLoS ONE, Vol 9, Iss 9, p e107981 (2014) |
| بيانات النشر: | Public Library of Science (PLoS), 2014. |
| سنة النشر: | 2014 |
| المجموعة: | LCC:Medicine LCC:Science |
| مصطلحات موضوعية: | Medicine, Science |
| الوصف: | DNA phosphorothioate (PT) modification is a recently identified epigenetic modification that occurs in the sugar-phosphate backbone of prokaryotic DNA. Previous studies have demonstrated that DNA PT modification is governed by the five DndABCDE proteins in a sequence-selective and RP stereo-specific manner. Bacteria may have acquired this physiological modification along with dndFGH as a restriction-modification system. However, little is known about the biological function of Dnd proteins, especially the smallest protein, DndE, in the PT modification pathway. DndE was reported to be a DNA-binding protein with a preference for nicked dsDNA in vitro; the binding of DndE to DNA occurs via six positively charged lysine residues on its surface. The substitution of these key lysine residues significantly decreased the DNA binding affinities of DndE proteins to undetectable levels. In this study, we conducted site-directed mutagenesis of dndE on a plasmid and measured DNA PT modifications under physiological conditions by mass spectrometry. We observed distinctive differences from the in vitro binding assays. Several mutants with lysine residues mutated to alanine decreased the total frequency of PT modifications, but none of the mutants completely eliminated PT modification. Our results suggest that the nicked dsDNA-binding capacity of DndE may not be crucial for PT modification and/or that DndE may have other biological functions in addition to binding to dsDNA. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1932-6203 24486930 |
| Relation: | http://europepmc.org/articles/PMC4182426?pdf=render; https://doaj.org/toc/1932-6203 |
| DOI: | 10.1371/journal.pone.0107981 |
| URL الوصول: | https://doaj.org/article/e057abf21ac244869309e4684d6dc090 |
| رقم الأكسشن: | edsdoj.057abf21ac244869309e4684d6dc090 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 19326203 24486930 |
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| DOI: | 10.1371/journal.pone.0107981 |