title: The Dnmt3L ADD Domain Controls Cytosine Methylation Establishment during Spermatogenesis.
abstract: A critical aspect of mammalian gametogenesis is the reprogramming of genomic DNA methylation. The catalytically inactive adaptor Dnmt3L is essential to ensuring this occurs correctly, but the mechanism by which it functions is unclear. Using gene targeting to engineer a single-amino-acid mutation, we show that the Dnmt3L histone H3 binding domain (ADD) is necessary for spermatogenesis. Genome-wide single-base-resolution DNA methylome analysis of mutant germ cells revealed overall reductions in CG methylation at repetitive sequences and non-promoter CpG islands. Strikingly, we also observe an even more severe loss of non-CG methylation, suggesting an unexpected role for the ADD in this process. These epigenetic deficiencies were coupled with defects in spermatogonia, with mutant cells displaying marked changes in gene expression and reactivation of retrotransposons. Our results demonstrate that the Dnmt3L ADD is necessary for Dnmt3L function and full reproductive fitness.
authors: Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors: Georgios
authors: G
authors: Vlachogiannis
authors: Department of Genetics, University of Georgia, 120 East Green Street, Athens, GA 30602, USA.
authors: Chad E
authors: CE
authors: Niederhuth
authors: Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors: Salih
authors: S
authors: Tuna
authors: Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors: Athanasia
authors: A
authors: Stathopoulou
authors: Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors: Keijo
authors: K
authors: Viiri
authors: Center for Reproductive Medicine, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, the Netherlands.
authors: Dirk G
authors: DG
authors: de Rooij
authors: Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors: Richard G
authors: RG
authors: Jenner
authors: Department of Genetics, University of Georgia, 120 East Green Street, Athens, GA 30602, USA.
authors: Robert J
authors: RJ
authors: Schmitz
authors: Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK. Electronic address: s.ooi@ucl.ac.uk.
authors: Steen K T
authors: SKT
authors: Ooi