Chromatin Targeting of de Novo DNA Methyltransferases by the PWWP Domain

Ying-Zi Ge,Min-Tie Pu,Humaira Gowher,Hai-Ping Wu,Jian-Ping Ding,Albert Jeltsch,Guo-Liang Xu

doi:10.1074/jbc.m312296200

Abstract

DNA methylation patterns of mammalian genomes are generated in gametogenesis and early embryonic development. Two de novo DNA methyltransferases, Dnmt3a and Dnmt3b, are responsible for the process. Both enzymes contain a long N-terminal regulatory region linked to a conserved C-terminal domain responsible for the catalytic activity. Although a PWWP domain in the N-terminal region has been shown to bind DNA in vitro, it is unclear how the DNA methyltransferases access their substrate in chromatin in vivo. We show here that the two proteins are associated with chromatin including mitotic chromosomes in mammalian cells, and the PWWP domain is essential for the chromatin targeting of the enzymes. The functional significance of PWWP-mediated chromatin targeting is suggested by the fact that a missense mutation in this domain of human DNMT3B causes immunodeficiency, centromeric heterochromatin instability, facial anomalies (ICF) syndrome, which is characterized by loss of methylation in satellite DNA, pericentromeric instability, and immunodeficiency. We demonstrate that the mutant protein completely loses its chromatin targeting capacity. Our data establish the PWWP domain as a novel chromatin/chromosome-targeting module and suggest that the PWWP-mediated chromatin association is essential for the function of the de novo methyltransferases during development.

Highlights

DNA methylation patterns of mammalian genomes are generated in gametogenesis and early embryonic development
We show here that the two proteins are associated with chromatin including mitotic chromosomes in mammalian cells, and the PWWP domain is essential for the chromatin targeting of the enzymes
The functional significance of PWWPmediated chromatin targeting is suggested by the fact that a missense mutation in this domain of human DNMT3B causes immunodeficiency, centromeric heterochromatin instability, facial anomalies (ICF) syndrome, which is characterized by loss of methylation in satellite DNA, pericentromeric instability, and immunodeficiency

Summary

Chromatin Targeting of de Novo DNA Methyltransferases by the PWWP Domain*

Two de novo DNA methyltransferases, Dnmt3a and Dnmt3b, are responsible for the process Both enzymes contain a long N-terminal regulatory region linked to a conserved C-terminal domain responsible for the catalytic activity. The functional significance of PWWPmediated chromatin targeting is suggested by the fact that a missense mutation in this domain of human DNMT3B causes immunodeficiency, centromeric heterochromatin instability, facial anomalies (ICF) syndrome, which is characterized by loss of methylation in satellite DNA, pericentromeric instability, and immunodeficiency. The generation of somatic methylation patterns in gametogenesis and embryogenesis depends on the de novo methyltransferases Dnmt3a and Dnmt3b [8, 9] Disruption of both or either of the genes in mice led to failure in methylation initiation in early embryos and caused lethal phenotypes [10]. Our findings underscore the importance of PWWP-mediated chromatin targeting for the function of DNA methyltransferases and provide a framework for future study of regulation of DNA methylation in development and pathogenesis

EXPERIMENTAL PROCEDURES

RESULTS

DISCUSSION