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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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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Sex-specific promoters regulate Dnmt3L expression in mouse germ cells

T.C. Shovlin1,4, D. Bourc’his2,5, S. La Salle3, A. O’Doherty1, J.M. Trasler3, T.H. Bestor2 and C.P. Walsh1,6

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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