| RRC ID |
54664
|
| Author |
Chatterjee B, Lin MH, Chen CC, Peng KL, Wu MS, Tseng MC, Chen YJ, Shen CJ.
|
| Title |
DNA Demethylation by DNMT3A and DNMT3B in vitro and of Methylated Episomal DNA in Transiently Transfected Cells.
|
| Journal |
Biochim Biophys Acta Gene Regul Mech
|
| Abstract |
The DNA methylation program in vertebrates is an essential part of the epigenetic regulatory cascade of development, cell differentiation, and progression of diseases including cancer. While the DNA methyltransferases (DNMTs) are responsible for the in vivo conversion of cytosine (C) to methylated cytosine (5mC), demethylation of 5mC on cellular DNA could be accomplished by the combined action of the ten-eleven translocation (TET) enzymes and DNA repair. Surprisingly, the mammalian DNMTs also possess active DNA demethylation activity in vitro in a Ca2+- and redox conditions-dependent manner, although little is known about its molecular mechanisms and occurrence in a cellular context. In this study, we have used LC-MS/MS to track down the fate of the methyl group removed from 5mC on DNA by mouse DNMT3B in vitro and found that it becomes covalently linked to the DNA methylation catalytic cysteine of the enzyme. We also show that Ca2+ homeostasis-dependent but TET1/TET2/TET3/TDG-independent demethylation of methylated episomal DNA by mouse DNMT3A or DNMT3B can occur in transfected human HEK 293 and mouse embryonic stem (ES) cells. Based on these results, we present a tentative working model of Ca2+ and redox conditions-dependent active DNA demethylation by DNMTs. Our study substantiates the potential roles of the vertebrate DNMTs as double-edged swords in DNA methylation-demethylation during Ca2+-dependent physiological processes.
|
| Volume |
1861(11)
|
| Pages |
1048-1061
|
| Published |
2018-11-1
|
| DOI |
10.1016/j.bbagrm.2018.09.009
|
| PII |
S1874-9399(18)30186-X
|
| PMID |
30300721
|
| MeSH |
Animals
Cell Line
DNA / metabolism
DNA (Cytosine-5-)-Methyltransferases / metabolism*
DNA Demethylation*
DNA Methyltransferase 3A
DNA-Binding Proteins / physiology
Humans
Mice
Mouse Embryonic Stem Cells / metabolism
Plasmids
Proto-Oncogene Proteins / physiology
Transfection
|
| IF |
5.179
|
| Times Cited |
1
|
| Resource |
| Human and Animal Cells |
Dnmt1-/-Dnmt3a-/-Dnmt3b-/- ES (clone19)(AES0146) |
