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Identification of Novel Imprinted Genes in a Genome-Wide Screen for Maternal Methylation

Developmental Genetics Program, The Babraham Institute, Cambridge CB2 4AT, UK

A characteristic of imprinted genes is that the maternal and paternal alleles show differences in methylation. To perform a genome-wide screen for novel imprinted loci, we applied methylation-sensitive representational difference analysis (Me-RDA) to parthenogenetic mouse embryos, to identify differentially methylated regions (DMRs) methylated specifically on the maternal allele. We isolated a total of 26 distinct clones from known and novel DMRs and identified three novel imprinted genes. Nap1l5 is located on proximal chromosome 6 and encodes a protein with homology with nucleosome assembly proteins (NAPs); it has tissue-specific imprinting with expression from the paternal allele. We identified two DMRs on chromosome 15, a chromosome that was not thought to contain imprinted loci, and demonstrated that each is associated with a paternally expressed transcript. Peg13 gives rise to a noncoding RNA that is highly expressed in the brain and imprinted in all tissues examined. A DMR was also identified at the chromosome 15 Slc38a4 gene, which encodes a system A amino acid transporter; we show that Slc38a4 is imprinted in a tissue-specific manner. Interestingly, two of the three novel genes identified in this screen are located within the introns of other genes; their identification indicates that such "microimprinted" domains may be more common than previously thought.

[The sequence data from this study have been submitted to GenBank under accession nos. AY151252 and AY151253. The following individuals kindly provided reagents, samples, or unpublished information as indicated in the paper: C. Beechey, J. Peters and D. Bodle.]

E-MAIL gavin.kelsey{at}bbsrc.ac.uk; FAX 44-1223-496022.

Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.781503.

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