title: MicroRNA 665 Regulates Dentinogenesis through MicroRNA-Mediated Silencing and Epigenetic Mechanisms.
abstract: Studies of proteins involved in microRNA (miRNA) processing, maturation, and silencing have indicated the importance of miRNAs in skeletogenesis, but the specific miRNAs involved in this process are incompletely defined. Here, we identified miRNA 665 (miR-665) as a potential repressor of odontoblast maturation. Studies with cultured cell lines and primary embryonic cells showed that miR-665 represses the expression of early and late odontoblast marker genes and stage-specific proteases involved in dentin maturation. Notably, miR-665 directly targeted Dlx3 mRNA and decreased Dlx3 expression. Furthermore, RNA-induced silencing complex (RISC) immunoprecipitation and biotin-labeled miR-665 pulldown studies identified Kat6a as another potential target of miR-665. KAT6A interacted physically and functionally with RUNX2, activating tissue-specific promoter activity and prompting odontoblast differentiation. Overexpression of miR-665 reduced the recruitment of KAT6A to Dspp and Dmp1 promoters and prevented KAT6A-induced chromatin remodeling, repressing gene transcription. Taken together, our results provide novel molecular evidence that miR-665 functions in an miRNA-epigenetic regulatory network to control dentinogenesis.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Hannah M
authors.initials: HM
authors.lastname: Heair
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Austin G
authors.initials: AG
authors.lastname: Kemper
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Bhaskar
authors.initials: B
authors.lastname: Roy
authors.affiliation: Cell Culture Laboratory, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Helena B
authors.initials: HB
authors.lastname: Lopes
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Harunur
authors.initials: H
authors.lastname: Rashid
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: Clarke
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Lubana K
authors.initials: LK
authors.lastname: Afreen
authors.affiliation: Cell Culture Laboratory, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Emanuela P
authors.initials: EP
authors.lastname: Ferraz
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Eddy
authors.initials: E
authors.lastname: Kim
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Amjad
authors.initials: A
authors.lastname: Javed
authors.affiliation: Cell Culture Laboratory, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Marcio M
authors.initials: MM
authors.lastname: Beloti
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Mary
authors.initials: M
authors.lastname: MacDougall
authors.affiliation: Department of Oral and Maxillofacial Surgery, Institute of Oral Health Research, School of Dentistry, University of Alabama at Birmingham, Birmingham, Alabama, USA hassank@uab.edu.
authors.forename: Mohammad Q
authors.initials: MQ
authors.lastname: Hassan
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002843:Chromatin / Q000235:genetics; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D003804:Dentin / Q000196:embryology*; D003810:Dentinogenesis / Q000235:genetics*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D020868:Gene Silencing; D057809:HEK293 Cells; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D009804:Odontoblasts / Q000166:cytology*; D010447:Peptide Hydrolases / Q000096:biosynthesis; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D012795:Sialoglycoproteins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D000099311:Dentin Sialophosphoprotein
chemical_list: D002843:Chromatin; D050659:Core Binding Factor Alpha 1 Subunit; D016326:Extracellular Matrix Proteins; D051548:Histone Acetyltransferases; D018398:Homeodomain Proteins; D035683:MicroRNAs; D010447:Peptide Hydrolases; D010750:Phosphoproteins; D012795:Sialoglycoproteins; D014157:Transcription Factors; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C088334:Distal-less homeobox proteins; C000592932:MIRN665 microRNA, human; C493709:RUNX2 protein, human; C108712:KAT6A protein, human