title: O-Linked N-Acetylglucosamine (O-GlcNAc) Transferase and O-GlcNAcase Interact with Mi2β Protein at the Aγ-Globin Promoter.
abstract: One mode of γ-globin gene silencing involves a GATA-1·FOG-1·Mi2β repressor complex that binds to the -566 GATA site relative to the (A)γ-globin gene cap site. However, the mechanism of how this repressor complex is assembled at the -566 GATA site is unknown. In this study, we demonstrate that the O-linked N-acetylglucosamine (O-GlcNAc) processing enzymes, O-GlcNAc-transferase (OGT) and O-GlcNAcase (OGA), interact with the (A)γ-globin promoter at the -566 GATA repressor site; however, mutation of the GATA site to GAGA significantly reduces OGT and OGA promoter interactions in β-globin locus yeast artificial chromosome (β-YAC) bone marrow cells. When WT β-YAC bone marrow cells are treated with the OGA inhibitor Thiamet-G, the occupancy of OGT, OGA, and Mi2β at the (A)γ-globin promoter is increased. In addition, OGT and Mi2β recruitment is increased at the (A)γ-globin promoter when γ-globin becomes repressed in postconception day E18 human β-YAC transgenic mouse fetal liver. Furthermore, we show that Mi2β is modified with O-GlcNAc, and both OGT and OGA interact with Mi2β, GATA-1, and FOG-1. Taken together, our data suggest that O-GlcNAcylation is a novel mechanism of γ-globin gene regulation mediated by modulating the assembly of the GATA-1·FOG-1·Mi2β repressor complex at the -566 GATA motif within the promoter.
authors.affiliation: From the Department of Biochemistry and Molecular Biology.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: IntelligeneDx Cancer Genomics, Olathe, Kansas 66061.
authors.forename: Flávia C
authors.initials: FC
authors.lastname: Costa
authors.affiliation: From the Department of Biochemistry and Molecular Biology.
authors.forename: Ee Phie
authors.initials: EP
authors.lastname: Tan
authors.affiliation: From the Department of Biochemistry and Molecular Biology.
authors.forename: Nathan
authors.initials: N
authors.lastname: Bushue
authors.affiliation: Pharmacology, Toxicology and Therapeutics.
authors.forename: Luciano
authors.initials: L
authors.lastname: DiTacchio
authors.affiliation: Department of Biochemistry and Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, Massachusetts 02118, and.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Costello
authors.affiliation: Department of Biochemistry and Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, Massachusetts 02118, and.
authors.forename: Mark E
authors.initials: ME
authors.lastname: McComb
authors.affiliation: Department of Biochemistry and Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, Massachusetts 02118, and.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Whelan
authors.affiliation: From the Department of Biochemistry and Molecular Biology, Anatomy and Cell Biology, and Cancer Center, Institute for Reproductive Health and Regenerative Medicine, and kpeterson@kumc.edu.
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Peterson
authors.affiliation: From the Department of Biochemistry and Molecular Biology, Cancer Center, Institute for Reproductive Health and Regenerative Medicine, and Alzheimer's Disease Center, University of Kansas Medical Center, Kansas City, Kansas 66160, cslawson@kumc.edu.
authors.forename: Chad
authors.initials: C
authors.lastname: Slawson
mesh_terms: D000818:Animals; D050982:GATA1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D020014:K562 Cells; D051379:Mice; D008822:Mice, Transgenic; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D020218:Response Elements; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics / Q000378:metabolism*; D055545:gamma-Globins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C494689:Gata1 protein, mouse; D009687:Nuclear Proteins; D014157:Transcription Factors; C498432:ZFPM1 protein, human; C107310:Zfpm1 protein, mouse; D055545:gamma-Globins; D017351:N-Acetylglucosaminyltransferases; C449880:O-GlcNAc transferase; C110628:hexosaminidase C; D001619:beta-N-Acetylhexosaminidases
keywords: CHD4; FOG; GATA transcription factor; Globin; O-GlcNAc-transferase; O-GlcNAcase; O-GlcNAcylation; O-linked N-acetylglucosamine (O-GlcNAc); post-translational modification (PTM); transcription