Cucurbit[7]uril-Driven Host–Guest Chemistry for Reversible Intervention of 5-Formylcytosine-Targeted Biochemical Reactions

Journal of the American Chemical Society

5 years ago

Cucurbit[7]uril-Driven Host–Guest Chemistry for Reversible Intervention of 5-Formylcytosine-Targeted Biochemical Reactions

Shao-Ru Wang, Xiang Zhou, Tian Tian, Lai Wei, Jia-Qi Wang, Yi-Nong Liu, Si-Min Liu, Yan-Yan Song, Xi-Ran Yang, Bo-Shi Fu, Chao-Xing Liu

5-Formylcytosine (5fC) is identified as one of the key players in active DNA demethylation and also as an epigenetic mark in mammals, thus representing a novel attractive target to chemical intervention. The current study represents an attempt to develop a reversible 5fC-targeted intervention tool. A supramolecular aldehyde reactive probe was therefore introduced for selective conversion of the 5fC to 5fC-AD nucleotide. Using various methods, we demonstrate that cucurbit[7]uril (CB7) selectively targets the 5fC-AD nucleotide in DNA, however, the binding of CB7 to 5fC-AD does not affect the hydrogen bonding properties of natural nucleobases in duplex DNA. Importantly, CB7-driven host–guest chemistry has been applied for reversible intervention of a variety of 5fC-targeted biochemical reactions, including restriction endonuclease digestion, DNA polymerase elongation, and polymerase chain reaction. On the basis of the current study, the macrocyclic CB7 creates obstructions that, through steric hindrance, prevent the enzyme from binding to the substrate, whereas the CB7/5fC-AD host–guest interactions can be reversed by treatment with adamantanamine. Moreover, fragment- and site-specific identification of 5fC modification in DNA has been accomplished without sequence restrictions. These findings thus show promising potential of host–guest chemistry for DNA/RNA epigenetics.

Publisher URL: http://dx.doi.org/10.1021/jacs.7b09635

DOI: 10.1021/jacs.7b09635