Maintenance of paternal methylation and repression of the imprinted H19 gene requires MBD3.
Paternal repression of the imprinted H19 gene is mediated by a differentially methylated domain (DMD) that is essential to imprinting of both H19 and the linked and oppositely imprinted Igf2 gene. The mechanisms by which paternal-specific methylation of the DMD survive the period of genome-wide demethylation in the early embryo and are subsequently used to govern imprinted expression are not known. Methyl-CpG binding (MBD) proteins are likely candidates to explain how these DMDs are recognized to silence the locus, because they preferentially bind methylated DNA and recruit repression complexes with histone deacetylase activity. MBD RNA and protein are found in preimplantation embryos, and chromatin immunoprecipitation shows that MBD3 is bound to the H19 DMD. To test a role for MBDs in imprinting, two independent RNAi-based strategies were used to deplete MBD3 in early mouse embryos, with the same results. In RNAi-treated blastocysts, paternal H19 expression was activated, supporting the hypothesis that MBD3, which is also a member of the Mi-2/NuRD complex, is required to repress the paternal H19 allele. RNAi-treated blastocysts also have reduced levels of the Mi-2/NuRD complex protein MTA-2, which suggests a role for the Mi-2/NuRD repressive complex in paternal-specific silencing at the H19 locus. Furthermore, DNA methylation was reduced at the H19 DMD when MBD3 protein was depleted. In contrast, expression and DNA methylation were not disrupted in preimplantation embryos for other imprinted genes. These results demonstrate new roles for MBD3 in maintaining imprinting control region DNA methylation and silencing the paternal H19 allele. Finally, MBD3-depleted preimplantation embryos have reduced cell numbers, suggesting a role for MBD3 in cell division.
Reese
Kimberly J
KJ
Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America.
Lin
Shu
S
Verona
Raluca I
RI
Schultz
Richard M
RM
Bartolomei
Marisa S
MS
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002455:Cell Division / Q000235:genetics; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015536:Down-Regulation; D047108:Embryonic Development / Q000235:genetics; D005220:Fathers; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018392:Genomic Imprinting; D006655:Histone Deacetylases / Q000378:metabolism; D008297:Male; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011485:Protein Binding; D034622:RNA Interference; D062085:RNA, Long Noncoding; D023121:RNA, Messenger, Stored / Q000378:metabolism; D022661:RNA, Untranslated / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D004268:DNA-Binding Proteins; C094335:H19 long non-coding RNA; C492013:Mbd3 protein, mouse; D062085:RNA, Long Noncoding; D023121:RNA, Messenger, Stored; D022661:RNA, Untranslated; D016601:RNA-Binding Proteins; D014157:Transcription Factors; D006655:Histone Deacetylases; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex
Dev Biol. 2002 Jan 1;241(1):172-82
11784103
Genes Dev. 2000 Aug 1;14(15):1908-19
10921905
EMBO J. 2002 Dec 2;21(23):6560-70
12456662
Cancer Res. 2003 Aug 1;63(15):4677-83
12907649
Biol Reprod. 2003 Sep;69(3):902-14
12748125
Annu Rev Cell Dev Biol. 2003;19:237-59
14570570
Gene Expr Patterns. 2003 Dec;3(6):697-702
14643676
Nature. 2003 Dec 18;426(6968):857-62
14661031
Science. 2004 Jan 9;303(5655):238-40
14716017
Development. 2004 Aug;131(15):3727-35
15240554
Development. 1987 Mar;99(3):371-82
3653008
Cell. 1991 Feb 22;64(4):849-59
1997210
Nature. 1991 May 9;351(6322):153-5
1709450
Development. 1991 Sep;113(1):119-27
1764989
Nature. 1993 Nov 25;366(6453):362-5
8247133
Mol Reprod Dev. 1994 Feb;37(2):121-9
7545925
Nat Genet. 1995 Apr;9(4):407-13
7795647
Mol Reprod Dev. 1995 Jun;41(2):232-8
7654376
Genes Dev. 1995 Sep 1;9(17):2079-89
7544754
Nature. 2000 Feb 3;403(6769):501-2
10676950
Curr Biol. 2000 Apr 20;10(8):475-8
10801417
Nature. 2000 May 25;405(6785):482-5
10839546
Nature. 2000 May 25;405(6785):486-9
10839547
Biol Reprod. 2000 Jun;62(6):1526-35
10819752
Curr Biol. 2000 May 18;10(10):607-10
10837224
Curr Biol. 2000 Jul 13;10(14):853-6
10899010
Development. 2007 Mar;134(6):1123-32
17287250
Genes Cells. 2000 Aug;5(8):677-88
10947852
Genes Dev. 2001 Mar 15;15(6):710-23
11274056
Biochem Biophys Res Commun. 2001 Oct 12;287(5):1099-104
11587535
Hum Mol Genet. 2001 Dec 15;10(26):2983-7
11751680
Mol Cell Biol. 1997 Jan;17(1):309-17
8972211
Mol Cell Biol. 1997 Aug;17(8):4322-9
9234689
Genes Dev. 1998 Jul 15;12(14):2108-13
9679055
Mol Cell Biol. 1998 Nov;18(11):6538-47
9774669
Genes Dev. 1998 Dec 1;12(23):3693-702
9851976
Hum Mol Genet. 2005 Apr 15;14 Spec No 1:R113-20
15809263
Am J Med Genet C Semin Med Genet. 2005 Aug 15;137C(1):12-23
16010676
Cold Spring Harb Symp Quant Biol. 2004;69:113-8
16117639
Nat Cell Biol. 2006 Mar;8(3):285-92
16462733
Cytogenet Genome Res. 2006;113(1-4):153-8
16575175
Mol Cell Biol. 2006 Dec;26(24):9471-83
17030610
EMBO J. 2007 Feb 7;26(3):720-9
17255950
J Biol Chem. 2002 Sep 20;277(38):35434-9
12124384