title: Oct4/Sox2 binding sites contribute to maintaining hypomethylation of the maternal igf2/h19 imprinting control region.
abstract: A central question in genomic imprinting is how parental-specific DNA methylation of imprinting control regions (ICR) is established during gametogenesis and maintained after fertilization. At the imprinted Igf2/H19 locus, CTCF binding maintains the unmethylated state of the maternal ICR after the blastocyst stage. In addition, evidence from Beckwith-Wiedemann patients and cultured mouse cells suggests that two Sox-Oct binding motifs within the Igf2/H19 ICR also participate in maintaining hypomethylation of the maternal allele. We found that the Sox and octamer elements from both Sox-Oct motifs were required to drive hypomethylation of integrated transgenes in mouse embryonic carcinoma cells. Oct4 and Sox2 showed cooperative binding to the Sox-Oct motifs, and both were present at the endogenous ICR. Using a mouse with mutations in the Oct4 binding sites, we found that maternally transmitted mutant ICRs acquired partial methylation in somatic tissues, but there was little effect on imprinted expression of H19 and Igf2. A subset of mature oocytes also showed partial methylation of the mutant ICR, which suggested that the Sox-Oct motifs provide some protection from methylation during oogenesis. The Sox-Oct motifs, however, were not required for erasure of paternal methylation in primordial germ cells, which indicated that the oocyte methylation was acquired post-natally. Maternally inherited mutant ICRs were unmethylated in blastocysts, which suggested that at least a portion of the methylation in somatic tissues occurred after implantation. These findings provide evidence that Sox-Oct motifs contribute to ICR hypomethylation in post-implantation embryos and maturing oocytes and link imprinted DNA methylation with key stem cell/germline transcription factors.
authors: Department of Cell and Developmental Biology, University of Illinois Urbana-Champaign, Urbana, Illinois, United States of America ; Biology Department, College of the Ozarks, Point Lookout, Missouri, United States of America.
authors: David L
authors: DL
authors: Zimmerman
authors: Craig S
authors: CS
authors: Boddy
authors: Christopher S
authors: CS
authors: Schoenherr
mesh_terms: D000483:Alleles; D000818:Animals; D001665:Binding Sites; D001755:Blastocyst / Q000378:metabolism; D000076246:CCCTC-Binding Factor; D019175:DNA Methylation / Q000235:genetics*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018392:Genomic Imprinting; D005854:Germ Cells / Q000378:metabolism; D007335:Insulin-Like Growth Factor II / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D009154:Mutation; D059372:Nucleotide Motifs / Q000235:genetics; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation / Q000235:genetics; D019076:Transgenes
chemical_list: D000076246:CCCTC-Binding Factor; C000619698:Ctcf protein, mouse; C094335:H19 long non-coding RNA; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D012097:Repressor Proteins; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D007335:Insulin-Like Growth Factor II