Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 11, Pages 6203-6210Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b01152
Keywords
-
Categories
Funding
- National Natural Science Foundation of China [21327006, 21435008, 21621064]
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDB14030200]
- Key Research Program of Frontier Sciences, CAS
Ask authors/readers for more resources
DNA N-6-methyl-2'-deoxyadenosine (6mdA) is an epigenetic modification in both eukaryotes and bacteria. Here we exploited stable isotope -labeled deoxynucleoside [N-15(5)]-2'-deoxyadenosine ([N-15(5)]-dA) as an initiation tracer and for the first time developed a metabolically differential tracing code for monitoring DNA 6mdA in human cells. We demonstrate that the initiation tracer [N-15(5)]-dA undergoes a specific and efficient adenine deamination reaction leading to the loss the exocyclic amine N-15, and further utilizes the purine salvage pathway to generate mainly both [N-15(4)]-dA and [N-15(4)]-2'-deoxyguanosine ([N-15(4)]-dG) in mammalian genomes. However, [N-15(5)]-dA is largely retained in the genomes of mycoplasmas, which are often found in cultured cells and experimental animals. Consequently, the methylation of dA generates 6mdA with a consistent coding pattern, with a predominance of [N-15(4)]-6mdA. Therefore, mammalian DNA 6mdA can be potentially discriminated from that generated by infecting mycoplasmas. Collectively, we show a promising approach for identification of authentic DNA 6mdA in human cells and determine if the human cells are contaminated with mycoplasmas.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available