Development of Methods Derived from Iodine-Induced Specific Cleavage for Identification and Quantitation of DNA Phosphorothioate Modifications
| Autor: | Jinli Li, Michael S. DeMott, Zixin Deng, Lingxin Kong, Peter C. Dedon, Yihua Sun, Ying Chen, Delin You, Tao Zheng, Bo Cao, Sucheng Zhu |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
DNA
Bacterial phosphorothioate (PT) modifications PT-IC-Seq ICDS lcsh:QR1-502 Phosphorothioate Oligonucleotides Computational biology Cleavage (embryo) Biochemistry Genome Deep sequencing Article iodine-induced cleavage (ICA) lcsh:Microbiology Phosphates 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine DNA Modification Escherichia coli Molecular Biology 030304 developmental biology 0303 health sciences Nuclease biology Genomics DNA biology.organism_classification chemistry biology.protein 030217 neurology & neurosurgery Bacteria Genome Bacterial Sulfur Iodine |
| Zdroj: | Biomolecules, Vol 10, Iss 1491, p 1491 (2020) Biomolecules Volume 10 Issue 11 |
| Popis: | DNA phosphorothioate (PT) modification is a novel modification that occurs on the DNA backbone, which refers to a non-bridging phosphate oxygen replaced by sulfur. This exclusive DNA modification widely distributes in bacteria but has not been found in eukaryotes to date. PT modification renders DNA nuclease tolerance and serves as a constitute element of bacterial restriction&ndash modification (R&ndash M) defensive system and more biological functions are awaiting exploration. Identification and quantification of the bacterial PT modifications are thus critical to better understanding their biological functions. This work describes three detailed methods derived from iodine-induced specific cleavage-an iodine-induced cleavage assay (ICA), a deep sequencing of iodine-induced cleavage at PT site (ICDS) and an iodine-induced cleavage PT sequencing (PT-IC-Seq)-for the investigation of PT modifications. Using these approaches, we have identified the presence of PT modifications and quantized the frequency of PT modifications in bacteria. These characterizations contributed to the high-resolution genomic mapping of PT modifications, in which the distribution of PT modification sites on the genome was marked accurately and the frequency of the specific modified sites was reliably obtained. Here, we provide time-saving and less labor-consuming methods for both of qualitative and quantitative analysis of genomic PT modifications. The application of these methodologies will offer great potential for better understanding the biology of the PT modifications and open the door to future further systematical study. |
| Databáze: | OpenAIRE |
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