title: Engineering of a Histone-Recognition Domain in Dnmt3a Alters the Epigenetic Landscape and Phenotypic Features of Mouse ESCs.
abstract: Histone modification and DNA methylation are associated with varying epigenetic "landscapes," but detailed mechanistic and functional links between the two remain unclear. Using the ATRX-DNMT3-DNMT3L (ADD) domain of the DNA methyltransferase Dnmt3a as a paradigm, we apply protein engineering to dissect the molecular interactions underlying the recruitment of this enzyme to specific regions of chromatin in mouse embryonic stem cells (ESCs). By rendering the ADD domain insensitive to histone modification, specifically H3K4 methylation or H3T3 phosphorylation, we demonstrate the consequence of dysregulated Dnmt3a binding and activity. Targeting of a Dnmt3a mutant to H3K4me3 promoters decreases gene expression in a subset of developmental genes and alters ESC differentiation, whereas aberrant binding of another mutant to H3T3ph during mitosis promotes chromosome instability. Our studies support the general view that histone modification "reading" and DNA methylation are closely coupled in mammalian cells, and suggest an avenue for the functional assessment of chromatin-associated proteins.
authors.affiliation: Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University, New York, NY 10065, USA.|European Molecular Biology Laboratory, Genome Biology Unit, 69117 Heidelberg, Germany.
authors.forename: Kyung-Min
authors.initials: KM
authors.lastname: Noh
authors.affiliation: Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing 100084, P.R. China.|MOE Key Laboratory of Protein Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, P.R. China.
authors.forename: Haibo
authors.initials: H
authors.lastname: Wang
authors.affiliation: Laboratory of RNA Molecular Biology, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Hyunjae R
authors.initials: HR
authors.lastname: Kim
authors.affiliation: Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Wenderski
authors.affiliation: Department of Medicine, Hematology-Oncology, Weill Cornell Medical College, New York, NY 10021, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Fang
authors.affiliation: Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Charles H
authors.initials: CH
authors.lastname: Li
authors.affiliation: Genomics, Resource Center, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Dewell
authors.affiliation: IV Drug Resistance Program, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Hughes
authors.affiliation: Department of Medicine, Hematology-Oncology, Weill Cornell Medical College, New York, NY 10021, USA.
authors.forename: Ari M
authors.initials: AM
authors.lastname: Melnick
authors.affiliation: Structure Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
authors.forename: Dinshaw J
authors.initials: DJ
authors.lastname: Patel
authors.affiliation: Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing 100084, P.R. China.|MOE Key Laboratory of Protein Sciences, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, P.R. China.|Collaborative Innovation Center for Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, P.R. China.
authors.forename: Haitao
authors.initials: H
authors.lastname: Li
authors.affiliation: Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University, New York, NY 10065, USA.
authors.forename: C David
authors.initials: CD
authors.lastname: Allis
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics*; D004265:DNA Helicases / Q000235:genetics; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D053595:Embryonic Stem Cells / Q000166:cytology*; D006657:Histones / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008938:Mitosis / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D000075924:X-linked Nuclear Protein
chemical_list: C000715535:Dnmt3a protein, mouse; D006657:Histones; D009687:Nuclear Proteins; C498796:Dnmt3l protein, mouse; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D004265:DNA Helicases; C490462:Atrx protein, mouse; D000075924:X-linked Nuclear Protein