Role of the Dnmt3 family in de novo methylation of imprinted and repetitive sequences during male germ cell development in the mouse.
DNA methylation is an important epigenetic modification regulating various biological phenomena, including genomic imprinting and transposon silencing. It is known that methylation of the differentially methylated regions (DMRs) associated with paternally imprinted genes and of some repetitive elements occurs during male germ cell development in the mouse. We have performed a detailed methylation analysis of the paternally methylated DMRs (H19, Dlk1/Gtl2 and Rasgrf1), interspersed repeats [SineB1, intracisternal A particle (IAP) and Line1] and satellite repeats (major and minor) to determine the timing of this de novo methylation in male germ cells. Furthermore, we have examined the roles of the de novo methyltransferases (Dnmt3a and Dnmt3b) and related protein (Dnmt3L) in this process. We found that methylation of all DMRs and repeats occurred progressively in fetal prospermatogonia and was completed by the newborn stage. Analysis of newborn prospermatogonia from germline-specific Dnmt3a and Dnmt3b knockout mice revealed that Dnmt3a mainly methylates the H19 and Dlk1/Gtl2 DMRs and a short interspersed repeat SineB1. Both Dnmt3a and Dnmt3b were involved in the methylation of Rasgrf1 DMR and long interspersed repeats IAP and Line1. Only Dnmt3b was required for the methylation of the satellite repeats. These results indicate both common and differential target specificities of Dnmt3a and Dnmt3b in vivo. Finally, all these sequences showed moderate to severe hypomethylation in Dnmt3L-deficient prospermatogonia, indicating the critical function and broad specificity of this factor in de novo methylation.
Kato
Yuzuru
Y
Department of Integrated Genetics, National Institute of Genetics, Mishima, Shizuoka, Japan.
Kaneda
Masahiro
M
Hata
Kenichiro
K
Kumaki
Kenji
K
Hisano
Mizue
M
Kohara
Yuji
Y
Okano
Masaki
M
Li
En
E
Nozaki
Masami
M
Sasaki
Hiroyuki
H
D000818:Animals; D000831:Animals, Newborn; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D004276:DNA, Satellite / Q000235:genetics / Q000378:metabolism; D005260:Female; D005333:Fetus / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000378:metabolism*; D020071:Interspersed Repetitive Sequences / Q000235:genetics; D008297:Male; D051379:Mice; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D013091:Spermatogenesis / Q000235:genetics; D000097584:DNA Methyltransferase 3B
D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D004276:DNA, Satellite; D000097584:DNA Methyltransferase 3B; C000715535:Dnmt3a protein, mouse; C498796:Dnmt3l protein, mouse