title: Binding of 14-3-3 reader proteins to phosphorylated DNMT1 facilitates aberrant DNA methylation and gene expression.
abstract: Mammalian DNA (cytosine-5) methyltransferase 1 (DNMT1) is essential for maintenance methylation. Phosphorylation of Ser143 (pSer143) stabilizes DNMT1 during DNA replication. Here, we show 14-3-3 is a reader protein of DNMT1pSer143. In mammalian cells 14-3-3 colocalizes and binds DNMT1pSer143 post-DNA replication. The level of DNMT1pSer143 increased with overexpression of 14-3-3 and decreased by its depletion. Binding of 14-3-3 proteins with DNMT1pSer143 resulted in inhibition of DNA methylation activity in vitro. In addition, overexpression of 14-3-3 in NIH3T3 cells led to decrease in DNMT1 specific activity resulting in hypomethylation of the genome that was rescued by transfection of DNMT1. Genes representing cell migration, mobility, proliferation and focal adhesion pathway were hypomethylated and overexpressed. Furthermore, overexpression of 14-3-3 also resulted in enhanced cell invasion. Analysis of TCGA breast cancer patient data showed significant correlation for DNA hypomethylation and reduced patient survival with increased 14-3-3 expressions. Therefore, we suggest that 14-3-3 is a crucial reader of DNMT1pSer143 that regulates DNA methylation and altered gene expression that contributes to cell invasion.
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Estève
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Zhang
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: V K Chaithanya
authors.initials: VK
authors.lastname: Ponnaluri
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Kanneganti
authors.initials: K
authors.lastname: Deepti
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Hang Gyeong
authors.initials: HG
authors.lastname: Chin
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Nan
authors.initials: N
authors.lastname: Dai
authors.affiliation: Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, TX 78957, USA.
authors.forename: Cari
authors.initials: C
authors.lastname: Sagum
authors.affiliation: Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, TX 78957, USA.
authors.forename: Karynne
authors.initials: K
authors.lastname: Black
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.
authors.forename: Ivan R
authors.initials: IR
authors.lastname: Corrêa
authors.affiliation: Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, TX 78957, USA.
authors.forename: Mark T
authors.initials: MT
authors.lastname: Bedford
authors.affiliation: Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Cheng
authors.affiliation: New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA pradhan@neb.com.
authors.forename: Sriharsa
authors.initials: S
authors.lastname: Pradhan
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics*; D004261:DNA Replication / Q000235:genetics; D005786:Gene Expression Regulation; D051379:Mice; D041681:NIH 3T3 Cells; D010766:Phosphorylation
chemical_list: D048948:14-3-3 Proteins; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619381:Dnmt1 protein, mouse