Webber A L, Tilghman S M
Howard Hughes Medical Institute and Department of Molecular Biology, Princeton University, New Jersey 08544, USA.
Mol Cell Biol. 1998 Apr;18(4):1903-10. doi: 10.1128/MCB.18.4.1903.
H19 and Igf2 are reciprocally imprinted genes that lie 90 kb apart on mouse chromosome 7. The two genes are coexpressed during development, with the H19 gene expressed exclusively from the maternal chromosome and Igf2 from the paternal chromosome. It has been proposed that their reciprocal imprinting is governed by a competition between the genes for a common set of enhancers. The competition on the paternal chromosome is influenced by extensive allele-specific methylation of the H19 gene and its 5' flank, which acts to inhibit H19 transcription and thus indirectly leads to the activation of the Igf2 gene. In contrast, no allele-specific methylation has been detected on the maternal chromosome, and the basis for the preference for H19 transcription on that chromosome is unresolved. In this investigation, the mechanism controlling the silencing of the Igf2 gene on the maternal chromosome was explored by studying the transcriptional activity of a yeast artificial chromosome (YAC) containing Igf2 and H19 following transfer into differentiated tissue culture cells. Contrary to expectations, both H19 and Igf2 were expressed from a single integrated copy of the YAC. Furthermore, Igf2 expression appeared to be independent of the H19 locus, based on deletions of the H19 gene promoter and its enhancers. These results suggest that an active process is responsible for the transcriptional bias toward H19 on the maternal chromosome and that the hypomethylated state of this chromosome cannot be viewed as a "default" state. Moreover, the active process is not reproduced in a differentiated cell and may require passage through the female germ line.
H19和Igf2是相互印记的基因,在小鼠7号染色体上相距90 kb。这两个基因在发育过程中共同表达,H19基因仅从母源染色体表达,Igf2从父源染色体表达。有人提出,它们的相互印记受这两个基因对一组共同增强子的竞争控制。父源染色体上的竞争受H19基因及其5'侧翼广泛的等位基因特异性甲基化影响,这种甲基化抑制H19转录,从而间接导致Igf2基因的激活。相反,在母源染色体上未检测到等位基因特异性甲基化,该染色体上H19转录偏好的基础仍未解决。在本研究中,通过研究转入分化的组织培养细胞后包含Igf2和H19的酵母人工染色体(YAC)的转录活性,探索了控制母源染色体上Igf2基因沉默的机制。与预期相反,H19和Igf2均从YAC的单个整合拷贝表达。此外,基于H19基因启动子及其增强子的缺失,Igf2的表达似乎独立于H19基因座。这些结果表明,一个活跃的过程导致了母源染色体上对H19的转录偏向,并且该染色体的低甲基化状态不能被视为“默认”状态。此外,这个活跃的过程在分化细胞中不会重现,可能需要通过雌性生殖系。