Distinct and overlapping DNMT1 interactions with multiple transcription factors in erythroid cells: Evidence for co-repressor functions.
DNMT1 is the maintenance DNA methyltransferase shown to be essential for embryonic development and cellular growth and differentiation in many somatic tissues in mammals. Increasing evidence has also suggested a role for DNMT1 in repressing gene expression through interactions with specific transcription factors. Previously, we identified DNMT1 as an interacting partner of the TR2/TR4 nuclear receptor heterodimer in erythroid cells, implicated in the developmental silencing of fetal β-type globin genes in the adult stage of human erythropoiesis. Here, we extended this work by using a biotinylation tagging approach to characterize DNMT1 protein complexes in mouse erythroleukemic cells. We identified novel DNMT1 interactions with several hematopoietic transcription factors with essential roles in erythroid differentiation, including GATA1, GFI-1b and FOG-1. We provide evidence for DNMT1 forming distinct protein subcomplexes with specific transcription factors and propose the existence of a "core" DNMT1 complex with the transcription factors ZBP-89 and ZNF143, which is also present in non-hematopoietic cells. Furthermore, we identified the short (17a.a.) PCNA Binding Domain (PBD) located near the N-terminus of DNMT1 as being necessary for mediating interactions with the transcription factors described herein. Lastly, we provide evidence for DNMT1 serving as a co-repressor of ZBP-89 and GATA1 acting through upstream regulatory elements of the PU.1 and GATA1 gene loci.
Division of Molecular Oncology, Biomedical Sciences Research Center "Alexander Fleming", Vari, Greece.
Dimitris N
DN
Papageorgiou
Division of Molecular Oncology, Biomedical Sciences Research Center "Alexander Fleming", Vari, Greece.
Elena
E
Karkoulia
Division of Molecular Oncology, Biomedical Sciences Research Center "Alexander Fleming", Vari, Greece.
Alexandra
A
Amaral-Psarris
Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
Pavel
P
Burda
Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands.
Katarzyna
K
Kolodziej
Proteomics Center, Erasmus Medical Center, Rotterdam, The Netherlands.
Jeroen
J
Demmers
Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL, USA.
Jörg
J
Bungert
Biocev, 1st Medical Faculty, Charles University, Prague, Czech Republic.
Tomas
T
Stopka
Division of Molecular Oncology, Biomedical Sciences Research Center "Alexander Fleming", Vari, Greece. Electronic address: john_strouboulis@imbb.forth.gr.
John
J
Strouboulis
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D041905:Erythroid Cells / Q000737:chemistry / Q000378:metabolism; D050982:GATA1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D051379:Mice; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D000099232:Proto-Oncogene Protein Spi-1
D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; D004268:DNA-Binding Proteins; D050982:GATA1 Transcription Factor; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D000099232:Proto-Oncogene Protein Spi-1; C494689:Gata1 protein, mouse; C486395:Gfi1b protein, mouse; C491741:Zfp148 protein, mouse; C107310:Zfpm1 protein, mouse; C000619380:DNMT1 protein, human; C000619381:Dnmt1 protein, mouse
DNMT1; Epigenetics; Erythropoiesis; Transcription factors