title: A methylation-stimulated DNA machine: an autonomous isothermal route to methyltransferase activity and inhibition analysis.
abstract: The operation of DNA nanomachines is generally triggered by either conformational changes of DNA nanostructure or external environmental stimuli. In the present study, we demonstrate an alternative driving force, DNA methylation, to stimulate DNA machine operation. DNA methylation changes neither DNA sequence and conformation nor external environment, however, blocks its cleavage by corresponding methylation-sensitive restriction endonuclease. We thus designed a strand displacement amplification DNA machine, which could be stimulated upon DNA methylation and then autonomously generates accumulated amounts of peroxidase-mimicking DNAzyme signaling machine products in an isothermal manner. The machine product DNAzyme could catalyze the H(2)O(2)-mediated oxidation of 2,2'-azino-bis(3-ethylbenzo thiazoline-6-sulfonic acid) (ABTS(2-)) to a colored product ABTS(·-). This methylation-stimulated DNA machine was further used as a colorimetric assay for analysis of methyltransferases activities and screening of methylation inhibitors. As compared with classical methylation assay, this facile isothermal DNA machine avoids the introduction of methylation-specific polymerase chain reaction and radioactive labels, which might be employed as an effective tool for DNA methylation analysis.
authors.affiliation: Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China.
authors.forename: Changfeng
authors.initials: C
authors.lastname: Zhu
authors.forename: Yanqin
authors.initials: Y
authors.lastname: Wen
authors.forename: Hongzhen
authors.initials: H
authors.lastname: Peng
authors.forename: Yitao
authors.initials: Y
authors.lastname: Long
authors.forename: Yao
authors.initials: Y
authors.lastname: He
authors.forename: Qing
authors.initials: Q
authors.lastname: Huang
authors.forename: Di
authors.initials: D
authors.lastname: Li
authors.forename: Chunhai
authors.initials: C
authors.lastname: Fan
mesh_terms: D004247:DNA / Q000737:chemistry*; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004795:Enzyme Stability; D007700:Kinetics
chemical_list: D004791:Enzyme Inhibitors; D004247:DNA; C029641:DNA modification methylase HpaII; D015257:DNA-Cytosine Methylases