A cis-acting control region is required exclusively for the tissue-specific imprinting of Gnas.
Genomic imprinting brings about allele-specific silencing according to parental origin. Silencing is controlled by cis-acting regulatory regions that are differentially marked during gametogenesis and can act over hundreds of kilobases to silence many genes. Two candidate imprinting control regions (ICRs) have been identified at the compact imprinted Gnas cluster on distal mouse chromosome 2, one at exon 1A upstream of Gnas itself and one covering the promoters for Gnasxl and the antisense Nespas (ref. 8). This imprinted cluster is complex, containing biallelic, maternally and paternally expressed transcripts that share exons. Gnas itself is mainly biallelically expressed but is weakly paternally repressed in specific tissues. Here we show that a paternally derived targeted deletion of the germline differentially methylated region at exon 1A abolishes tissue-specific imprinting of Gnas. This rescues the abnormal phenotype of mice with a maternally derived Gnas mutation. Imprinting of alternative transcripts, Nesp, Gnasxl and Nespas (ref. 13), in the cluster is unaffected. The results establish that the differentially methylated region at exon 1A contains an imprinting control element that specifically regulates Gnas and comprises a characterized ICR for a gene that is only weakly imprinted in a minority of tissues. There must be a second ICR regulating the alternative transcripts.
Williamson
Christine M
CM
MRC Mammalian Genetics Unit, Harwell, Oxfordshire, OX11 0RD, UK.
Ball
Simon T
ST
Nottingham
Wade T
WT
Skinner
Judith A
JA
Plagge
Antonius
A
Turner
Martin D
MD
Powles
Nicola
N
Hough
Tertius
T
Papworth
David
D
Fraser
William D
WD
Maconochie
Mark
M
Peters
Jo
J
D000818:Animals; D002864:Chromogranins; D019175:DNA Methylation; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics*; D018390:Gene Targeting; D018392:Genomic Imprinting; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009154:Mutation; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid
D002864:Chromogranins; C498642:Gnas protein, mouse; D019205:GTP-Binding Protein alpha Subunits, Gs