Development of an Endonuclease V-Assisted Analytical Method for Sequencing Analysis of Deoxyinosine in DNA.

Abstract

Deoxyinosine (dI) is a highly mutagenic lesion that preferentially pairs with deoxycytidine during replication, which may induce A to G transition and ultimately contribute to carcinogenesis. Therefore, finding the site of dI modification in DNA is of great value for both basic research and clinical applications. Herein, we developed a novel method to sequence the dI modification site in DNA, which utilizes endonuclease V (EndoV)-dependent deamination repair to specifically label the modification site with biotin-14-dATP that allows the affinity enrichment of dI-bearing DNA for sequencing. We have achieved efficient determination of the location of the modified nucleotide in dI-bearing plasmid DNA with the assistance of EndoV-dependent deamination repair. We have also successfully applied this approach to locate the dI modification sites in the mitochondrial DNA of human cells. Our method should be generally applicable for genome-wide sequencing analysis of dI modifications in living organisms.

• Authors: Xiaofang Zheng, Di Chen, Yingqi Zhao, Xiaoxia Dai, Changjun You
• Journal: Analytical chemistry (Anal Chem) Vol. 94 Issue 33 Pg. 11627-11632 (08 23 2022) ISSN: 1520-6882 [Electronic] United States
• PMID: 35942621 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Inosine
• DNA
• Deoxyribonuclease (Pyrimidine Dimer)
• deoxyinosine

Topics

• DNA (genetics)
• DNA Repair
• Deoxyribonuclease (Pyrimidine Dimer) (genetics, metabolism)
• Humans
• Inosine (analogs & derivatives)