| Summary: | <p>Abstract</p> <p>Background-</p> <p>Parent-specific methylation of specific CpG residues is critical to imprinting in eutherian mammals, but its importance to imprinting in marsupials and, thus, the evolutionary origins of the imprinting mechanism have been the subject of controversy. This has been particularly true for the imprinted <it>Insulin-like Growth Factor II </it>(<it>IGF2</it>), a key regulator of embryonic growth in vertebrates and a focal point of the selective forces leading to genomic imprinting. The presence of the essential imprinting effector, <it>DNMT3L</it>, in marsupial genomes and the demonstration of a differentially methylated region (DMR) in the retrotransposon-derived imprinted gene, <it>PEG10</it>, in tammar wallaby argue for a role for methylation in imprinting, but several studies have found no evidence of parent-specific methylation at other imprinted loci in marsupials.</p> <p>Results-</p> <p>We performed the most extensive search to date for allele-specific patterns of CpG methylation within CpG isochores or CpG enriched segments across a 22 kilobase region surrounding the <it>IGF2 </it>gene in the South American opossum <it>Monodelphis domestica</it>. We identified a previously unknown 5'-untranslated exon for opossum <it>IGF2</it>, which is flanked by sequences defining a putative neonatal promoter, a DMR and an active Matrix Attachment Region (MAR). Demethylation of this DMR in opossum neonatal fibroblasts results in abherrant biallelic expression of <it>IGF2</it>.</p> <p>Conclusion-</p> <p>The demonstration of a DMR and an active MAR in the 5' flank of opossum <it>IGF2 </it>mirrors the regulatory features of the 5' flank of <it>Igf2 </it>in mice. However, demethylation induced activation of the maternal allele of <it>IGF2 </it>in opossum differs from the demethylation induced repression of the paternal <it>Igf2 </it>allele in mice. While it can now be concluded that parent-specific DNA methylation is an epigentic mark common to Marsupialia and Eutheria, the molecular mechanisms of transcriptional silencing at imprinted loci have clearly evolved along independent trajectories.</p>
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