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Molecular and Cellular Biology, April 2002, p. 2089-2098, Vol. 22, No. 7
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.7.2089-2098.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Shared Role for Differentially Methylated Domains of Imprinted Genes

Bonnie Reinhart,1 Mariam Eljanne,2,{dagger} and J. Richard Chaillet3*

Department of Biological Sciences,1 Department of Human Genetics,2 Department of Pediatrics, University of Pittsburgh, Pittsburgh, Pennsylvania 152133

Received 26 September 2001/ Returned for modification 2 November 2001/ Accepted 4 January 2002

For most imprinted genes, a difference in expression between the maternal and paternal alleles is associated with a corresponding difference in DNA methylation that is localized to a differentially methylated domain (DMD). Removal of a gene's DMD leads to a loss of imprinting. These observations suggest that DMDs have a determinative role in genomic imprinting. To examine this possibility, we introduced sequences from the DMDs of the imprinted Igf2r, H19, and Snrpn genes into a nonimprinted derivative of the normally imprinted RSVIgmyc transgene, created by excising its own DMD. Hybrid transgenes with sequences from the Igf2r DMD2 were consistently imprinted, with the maternal allele being more methylated than the paternal allele. Only the repeated sequences within DMD2 were required for imprinting these transgenes. Hybrid transgenes containing H19 and Snrpn DMD sequences and ones containing sequences from the long terminal repeat of a murine intracisternal A particle retrotransposon were not imprinted. The Igf2r hybrid transgenes are comprised entirely of mouse genomic DNA and behave as endogenous imprinted genes in inbred wild-type and mutant mouse strains. These types of hybrid transgenes can be used to elucidate the functions of DMD sequences in genomic imprinting.

* Corresponding author. Mailing address: Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA 15213. Phone: (412) 692-7993. Fax: (412) 692-6096. E-mail: Chaillet{at}pitt.edu.

{dagger} Present address: Cytyc Corporation, Boxborough, MA 01719.

Molecular and Cellular Biology, April 2002, p. 2089-2098, Vol. 22, No. 7
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.7.2089-2098.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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