Dnmt2 is not required for de novo and maintenance methylation of viral DNA in embryonic stem cells.
We have shown previously that de novo methylation activities persist in mouse embryonic stem (ES) cells homozygous for a null mutation of Dnmt1 that encodes the major DNA cytosine methyltransferase. In this study, we have cloned a putative mammalian DNA methyltransferase gene, termed Dnmt2 , that is homologous to pmt1 of fission yeast. Different from pmt1 in which the catalytic Pro-Pro-Cys (PPC) motif is 'mutated' to Pro-Ser-Cys, Dnmt2 contains all the conserved methyltransferase motifs, thus likely encoding a functional cytosine methyltransferase. However, baculovirus-expressed Dnmt2 protein failed to methylate DNA in vitro . To investigate whether Dnmt2 functions as a DNA methyltransferase in vivo , we inactivated the Dnmt2 gene by targeted deletion of the putative catalytic PPC motif in ES cells. We showed that endogenous virus was fully methylated in Dnmt2 -deficient mutant ES cells. Furthermore, newly integrated retrovirus DNA was methylated de novo in infected mutant ES cells as efficiently as in wild-type cells. These results indicate that Dnmt2 is not essential for global de novo or maintenance methylation of DNA in ES cells.
Okano
M
M
Cardiovascular Research Center, Massachusetts General Hospital, Department of Medicine,Harvard Medical School, 149, 13th Street, Charlestown, MA 02129, USA.
Xie
S
S
Li
E
E
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D003001:Cloning, Molecular; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D004279:DNA, Viral / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics; D011533:Proviruses / Q000235:genetics; D012568:Schizosaccharomyces; D013234:Stem Cells
D004279:DNA, Viral; C491786:Dnmt2 protein, mouse; D004248:DNA (Cytosine-5-)-Methyltransferases; C112218:TRDMT1 protein, human