Genomic imprinting of IGF2 in marsupials is methylation dependent.
BACKGROUND
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 Insulin-like Growth Factor II (IGF2), 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, DNMT3L, in marsupial genomes and the demonstration of a differentially methylated region (DMR) in the retrotransposon-derived imprinted gene, PEG10, 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.


RESULTS
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 IGF2 gene in the South American opossum Monodelphis domestica. We identified a previously unknown 5'-untranslated exon for opossum IGF2, 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 IGF2.


CONCLUSIONS
The demonstration of a DMR and an active MAR in the 5' flank of opossum IGF2 mirrors the regulatory features of the 5' flank of Igf2 in mice. However, demethylation induced activation of the maternal allele of IGF2 in opossum differs from the demethylation induced repression of the paternal Igf2 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.
Lawton
Betty R
BR
Department of Molecular and Cell Biology, University of Connecticut, 354 Mansfield Rd., Storrs, CT 06269, USA . betty.lawton@uconn.edu
Carone
Benjamin R
BR
Obergfell
Craig J
CJ
Ferreri
Gianni C
GC
Gondolphi
Christina M
CM
Vandeberg
John L
JL
Imumorin
Ikhide
I
O'Neill
Rachel J
RJ
O'Neill
Michael J
MJ
D000818:Animals; D005075:Biological Evolution; D004248:DNA (Cytosine-5-)-Methyltransferases; D019175:DNA Methylation; D018392:Genomic Imprinting; D040582:Inheritance Patterns; D007335:Insulin-Like Growth Factor II / Q000235:genetics*; D008394:Marsupialia / Q000235:genetics; D009893:Opossums / Q000235:genetics*
D007335:Insulin-Like Growth Factor II; D004248:DNA (Cytosine-5-)-Methyltransferases