Hum. Reprod. Advance Access originally published online on October 23, 2006
Human Reproduction 2007 22(2):457-467; doi:10.1093/humrep/del379
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Sex-specific promoters regulate Dnmt3L expression in mouse germ cells
1 Stem Cells and Epigenetics Research Group, Centre for Molecular Biosciences, School of Biomedical Sciences, University of Ulster, Coleraine, UK 2 Department of Genetics and Development, College of Physicians and Surgeons of Columbia University, New York, NY, USA and 3 Montreal Children’s Hospital Research Institute and Departments of Pediatrics, Human Genetics, Pharmacology & Therapeutics, McGill University, Montreal, QC, Canada
4 Present address: The Wellcome Trust/Cancer Research UK Gurdon Institute, The Henry Wellcome Building of Cancer and Developmental Biology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK
5 Present address: INSERM U741, Institut Jacques Monod, 2 Place Jussieu, 75251 Paris, CEDEX 05, France
6 To whom correspondence should be addressed at: Stem Cells and Epigenetics Research Group, Centre for Molecular Biosciences, School of Biomedical Sciences, University of Ulster, Coleraine BT52 1SA, UK. E-mail: cp.walsh{at}ulster.ac.uk
BACKGROUND: Dnmt3L, a member of the DNA methyltransferase 3 family, lacks enzymatic activity but is required for de-novo methylation of imprinted genes in oocytes and for transposon repression in male germ cells. METHODS: We used northern blots, RT–PCR, 5' rapid amplification of complementary DNA (cDNA) ends (RACE), RNase H mapping, real-time/quantitative RT–PCR and in situ hybridization to identify and characterize Dnmt3L transcripts produced during germ cell development. RESULTS: Mouse Dnmt3L uses three sex-specific promoters, not the single promoter previously thought. A promoter active in prospermatogonia drives transcription of an mRNA encoding the full-length protein in perinatal testis, where de-novo methylation occurs. Late pachytene spermatocytes activate a second promoter in intron 9 of the Dnmt3L gene. After this stage, the predominant transcripts are three truncated mRNAs, which appear to be non-coding. We could also detect similar adult testis transcripts in humans. In the mouse ovary, an oocyte-specific promoter located in an intron of the neighbouring autoimmune regulator (Aire) gene produces a transcript with the full open reading frame (ORF). This is the only Dnmt3L transcript found in growing oocytes and is absent in the oocytes of Dnmt3L–/– females. CONCLUSIONS: Sex-specific promoters control Dnmt3L expression in the mouse germ line, mirroring the situation at the Dnmt1 and Dnmt3A loci.
Key words: Dnmt3L/imprinting/oocyte/promoter/testis
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