title: G9a and ZNF644 Physically Associate to Suppress Progenitor Gene Expression during Neurogenesis.
abstract: Proliferating progenitor cells undergo changes in competence to give rise to post-mitotic progeny of specialized function. These cell-fate transitions typically involve dynamic regulation of gene expression by histone methyltransferase (HMT) complexes. However, the composition, roles, and regulation of these assemblies in regulating cell-fate decisions in vivo are poorly understood. Using unbiased affinity purification and mass spectrometry, we identified the uncharacterized C2H2-like zinc finger protein ZNF644 as a G9a/GLP-interacting protein and co-regulator of histone methylation. In zebrafish, functional characterization of ZNF644 orthologs, znf644a and znf644b, revealed complementary roles in regulating G9a/H3K9me2-mediated gene silencing during neurogenesis. The non-overlapping requirements for znf644a and znf644b during retinal differentiation demarcate critical aspects of retinal differentiation programs regulated by differential G9a-ZNF644 associations, such as transitioning proliferating progenitor cells toward differentiation. Collectively, our data point to ZNF644 as a critical co-regulator of G9a/H3K9me2-mediated gene silencing during neuronal differentiation.
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, Medical Science Building, Toronto, ON M5S 3E1, Canada.
authors.forename: Jonathan B
authors.initials: JB
authors.lastname: Olsen
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, ON, M5S 3G5, Canada.
authors.forename: Loksum
authors.initials: L
authors.lastname: Wong
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, ON, M5S 3G5, Canada.
authors.forename: Steven
authors.initials: S
authors.lastname: Deimling
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, ON, M5S 3G5, Canada.
authors.forename: Amanda
authors.initials: A
authors.lastname: Miles
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Guo
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, Medical Science Building, Toronto, ON M5S 3E1, Canada; Department of Computer Science, University of Toronto, Toronto, ON M5S 3G4, Canada.
authors.forename: Yue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, Medical Science Building, Toronto, ON M5S 3E1, Canada; Department of Computer Science, University of Toronto, Toronto, ON M5S 3G4, Canada.
authors.forename: Zhaolei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, Medical Science Building, Toronto, ON M5S 3E1, Canada.
authors.forename: Jack F
authors.initials: JF
authors.lastname: Greenblatt
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, Canada; Department of Molecular Genetics, University of Toronto, Medical Science Building, Toronto, ON M5S 3E1, Canada. Electronic address: andrew.emili@utoronto.ca.
authors.forename: Andrew
authors.initials: A
authors.lastname: Emili
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, ON, M5S 3G5, Canada; Department of Ophthalmology and Vision Sciences, University of Toronto, Toronto, ON M5T 3A9, Canada; Centre for the Analysis of Genome Evolution and Function, University of Toronto, Toronto, ON M5S 3B2, Canada. Electronic address: v.tropepe@utoronto.ca.
authors.forename: Vincent
authors.initials: V
authors.lastname: Tropepe
mesh_terms: D000818:Animals; D001665:Binding Sites; D015415:Biomarkers; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D006649:Histocompatibility Antigens / Q000235:genetics / Q000378:metabolism*; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D008745:Methylation; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism*; D055495:Neurogenesis / Q000235:genetics*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D011485:Protein Binding; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D012160:Retina / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D015415:Biomarkers; D006649:Histocompatibility Antigens; D006657:Histones; D014157:Transcription Factors; C573234:ZNF644 protein, human; C445405:EHMT2 protein, human; D011495:Histone-Lysine N-Methyltransferase