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Molecular and Cellular Biology, August 2000, p. 5808-5817, Vol. 20, No. 16
0270-7306/00/$04.00+0

Identification of a Methylation Imprint Mark within the Mouse Gnas Locus

Jie Liu, Shuhua Yu, Deborah Litman, Weiping Chen, and Lee S. Weinstein*

Metabolic Diseases Branch, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892

Received 10 March 2000/Returned for modification 2 May 2000/Accepted 22 May 2000

The imprinted mouse gene Gnas produces the G protein alpha -subunit GSalpha and several other gene products by using alternative promoters and first exons. GSalpha is maternally expressed in some tissues and biallelically expressed in most other tissues, while the gene products NESP55 and XLalpha s are maternally and paternally expressed, respectively. We investigated the mechanisms of Gnas imprinting. The GSalpha promoter and first exon are not methylated on either allele. A further upstream region (approximately from positions -3400 to -939 relative to the GSalpha translational start site) is methylated only on the maternal allele in all adult somatic tissues and in early postimplantation development. Within this region lies a fourth promoter and first exon (exon 1A) that generates paternal-specific mRNAs of unknown function. Exon 1A and GSalpha mRNAs have similar expression patterns, making competition between their promoters unlikely. Differential methylation in this region is established during gametogenesis, being present in oocytes and absent in spermatozoa, and is maintained in preimplantation E3.5d blastocysts. Therefore, this region is a methylation imprint mark. In contrast, differential methylation of the NESP55 and XLalpha s promoter regions (Nesp and Gnasxl) is not established during gametogenesis. The methylation imprint mark that we identified may be important for the tissue-specific imprinting of GSalpha .


* Corresponding author. Mailing address: Metabolic Diseases Branch, NIDDK/NIH, Bldg. 10, Rm. 8C101, Bethesda, MD 20892-1752. Phone: (301) 402-2923. Fax: (301) 402-0374. E-mail: leew{at}amb.niddk.nih.gov.


Molecular and Cellular Biology, August 2000, p. 5808-5817, Vol. 20, No. 16
0270-7306/00/$04.00+0



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