1. Department of Genetics and Development, College of Physicians and Surgeons of Columbia University, New York, New York 10032, USA
2. Department of Biochemistry, Emory University, Atlanta, Georgia 30322, USA
3. Laboratory of Chromatin Biology, The Rockefeller University, New York, New York 10021, USA
4. Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA
5. Institute of Biotechnology, National Taiwan University, Taipei 106, Taiwan
6. These authors contributed equally to this work.

Correspondence to: Xiaodong Cheng²Timothy H. Bestor¹ Correspondence and requests for materials should be addressed to T.H.B. (Email: THB12@columbia.edu) or X.C. (Email: XCheng@emory.edu).

Abstract

Mammals use DNA methylation for the heritable silencing of retrotransposons and imprinted genes and for the inactivation of the X chromosome in females. The establishment of patterns of DNA methylation during gametogenesis depends in part on DNMT3L, an enzymatically inactive regulatory factor that is related in sequence to the DNA methyltransferases DNMT3A and DNMT3B^{1, 2}. The main proteins that interact in vivo with the product of an epitope-tagged allele of the endogenous Dnmt3L gene were identified by mass spectrometry as DNMT3A2, DNMT3B and the four core histones. Peptide interaction assays showed that DNMT3L specifically interacts with the extreme amino terminus of histone H3; this interaction was strongly inhibited by methylation at lysine 4 of histone H3 but was insensitive to modifications at other positions. Crystallographic studies of human DNMT3L showed that the protein has a carboxy-terminal methyltransferase-like domain and an N-terminal cysteine-rich domain. Cocrystallization of DNMT3L with the tail of histone H3 revealed that the tail bound to the cysteine-rich domain of DNMT3L, and substitution of key residues in the binding site eliminated the H3 tail–DNMT3L interaction. These data indicate that DNMT3L recognizes histone H3 tails that are unmethylated at lysine 4 and induces de novo DNA methylation by recruitment or activation of DNMT3A2.

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