H19 and Igf2 monoallelic expression is regulated in two distinct ways by a shared cis acting regulatory region upstream of H19.
H19 and Igf2 are expressed in a monoallelic fashion from the maternal and paternal chromosomes, respectively. A region upstream of H19 has been shown to regulate such imprinted expression of both genes in cis. We have taken advantage of a loxP/cre recombinase-based strategy to delete this region in mice in a conditional manner to determine the temporal requirement of the upstream region in initiating and maintaining the imprinted expression of H19 and Igf2. Analysis of allele-specific expression of H19 and Igf2 and DNA methylation at the H19 promoter demonstrates that this region controls the monoallelic expression of the two genes in different ways, suggesting that it harbors two functionally distinct regulatory elements. Continued presence of the region is required to silence maternal Igf2 in accordance with its proposed role as an insulator. However, it does not have a direct role in keeping the paternal H19 promoter silenced. Instead, on the paternal chromosome, the upstream element mediates epigenetic modifications of the H19 promoter region during development, leading to transcriptional silencing of H19. Thereafter, its presence is redundant for preventing transcription. Presently, this temporal requirement of the silencing element appears to be a unique cis activity in the mammalian system. However, it is likely that other cis-acting elements, positive and negative, have the ability to effect stable changes in the chromatin structure and are not constantly required to give signals to the transcriptional machinery.
Srivastava
M
M
Laboratory of Mammalian Genes and Development, National Institute of Child Health and Human Development, Bethesda, Maryland 20892, USA.
Hsieh
S
S
Grinberg
A
A
Williams-Simons
L
L
Huang
S P
SP
Pfeifer
K
K
D000483:Alleles; D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics; D002875:Chromosomes / Q000235:genetics; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D005260:Female; D005718:Gametogenesis / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D020868:Gene Silencing; D018392:Genomic Imprinting / Q000235:genetics*; D007335:Insulin-Like Growth Factor II / Q000235:genetics*; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008957:Models, Genetic; D009124:Muscle Proteins / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D062085:RNA, Long Noncoding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D022661:RNA, Untranslated; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D017384:Sequence Deletion / Q000235:genetics; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics; D014764:Viral Proteins
C094335:H19 long non-coding RNA; D009124:Muscle Proteins; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D022661:RNA, Untranslated; D014764:Viral Proteins; D007335:Insulin-Like Growth Factor II; C045073:Cre recombinase; D019426:Integrases