O-GlcNAc Homeostasis Controls Cell Fate Decisions During Hematopoiesis

The addition of O-GlcNAc (a single β-D-N-acetylglucosamine sugar at serine and threonine residues) by O-GlcNAc transferase (OGT) and removal by O-GlcNAcase (OGA) maintains homeostatic levels of O-GlcNAc. We investigated the role of O-GlcNAc homeostasis in hematopoiesis utilizing G1E-ER4 cells carrying a GATA-1 transcription factor fused to the estrogen receptor (GATA-1ER) that undergo erythropoiesis following the addition of β-estradiol (E2) and myeloid leukemia cells that differentiate into neutrophils in the presence of all-trans retinoic acid. During G1E-ER4 differentiation, a decrease in overall O-GlcNAc levels and an increase in GATA-1 interactions with OGT and OGA were observed. Transcriptome analysis on G1E-ER4 cells differentiated in the presence of Thiamet-G (TMG), an OGA inhibitor, identified expression changes in 433 GATA-1 target genes. Chromatin immunoprecipitation demonstrated that the occupancy of GATA-1, OGT, and OGA at Laptm5 gene GATA site was decreased with TMG. Myeloid leukemia cells showed a decline in O-GlcNAc levels during differentiation and TMG reduced the expression of genes involved in differentiation. Sustained treatment with TMG in G1E-ER4 cells prior to differentiation caused a reduction of hemoglobin positive cells during differentiation. Our results show that alterations in O-GlcNAc homeostasis disrupt transcriptional programs causing differentiation errors suggesting a vital role of O-GlcNAcylation in control of cell fate.

[1]  R. Swerdlow,et al.  Sustained O-GlcNAcylation reprograms mitochondrial function to regulate energy metabolism , 2017, The Journal of Biological Chemistry.

[2]  N. Hastings,et al.  Inhibition of O-GlcNAcase leads to elevation of O-GlcNAc tau and reduction of tauopathy and cerebrospinal fluid tau in rTg4510 mice , 2017, Molecular Neurodegeneration.

[3]  K. Zhao,et al.  O-GlcNAcase Is an RNA Polymerase II Elongation Factor Coupled to Pausing Factors SPT5 and TIF1β* , 2016, The Journal of Biological Chemistry.

[4]  M. Reginato,et al.  O-GlcNAcylation in Cancer Biology: Linking Metabolism and Signaling. , 2016, Journal of molecular biology.

[5]  Andrew D. Rouillard,et al.  The harmonizome: a collection of processed datasets gathered to serve and mine knowledge about genes and proteins , 2016, Database J. Biol. Databases Curation.

[6]  C. Slawson,et al.  O-Linked N-Acetylglucosamine (O-GlcNAc) Transferase and O-GlcNAcase Interact with Mi2β Protein at the Aγ-Globin Promoter* , 2016, The Journal of Biological Chemistry.

[7]  A. Burlingame,et al.  Human RNA Polymerase II Promoter Recruitment in Vitro Is Regulated by O-Linked N-Acetylglucosaminyltransferase (OGT)* , 2016, The Journal of Biological Chemistry.

[8]  Salam A. Assi,et al.  Dynamic Gene Regulatory Networks Drive Hematopoietic Specification and Differentiation , 2016, Developmental cell.

[9]  S. Fucharoen,et al.  Erythropoietin and IGF-1 signaling synchronize cell proliferation and maturation during erythropoiesis , 2015, Genes & development.

[10]  K. Kamemura,et al.  Global increase in O-linked N-acetylglucosamine modification promotes osteoblast differentiation. , 2015, Experimental cell research.

[11]  Jennifer J. Kohler,et al.  Enhanced Transfer of a Photocross-linking N-Acetylglucosamine (GlcNAc) Analog by an O-GlcNAc Transferase Mutant with Converted Substrate Specificity* , 2015, The Journal of Biological Chemistry.

[12]  Y. Loh,et al.  RING1B O-GlcNAcylation regulates gene targeting of polycomb repressive complex 1 in human embryonic stem cells. , 2015, Stem cell research.

[13]  J. Hanover,et al.  You are what you eat: O-linked N-acetylglucosamine in disease, development and epigenetics , 2015, Current opinion in clinical nutrition and metabolic care.

[14]  Jürg Müller,et al.  A critical perspective of the diverse roles of O-GlcNAc transferase in chromatin , 2015, Chromosoma.

[15]  C. Slawson,et al.  O-GlcNAcase Expression is Sensitive to Changes in O-GlcNAc Homeostasis , 2014, Front. Endocrinol..

[16]  G. Hart.  Three Decades of Research on O-GlcNAcylation – A Major Nutrient Sensor That Regulates Signaling, Transcription and Cellular Metabolism , 2014, Front. Endocrinol..

[17]  G. Barton,et al.  Elevated O‐GlcNAc Levels Activate Epigenetically Repressed Genes and Delay Mouse ESC Differentiation Without Affecting Naïve to Primed Cell Transition , 2014, Stem cells.

[18]  G. Hart,et al.  Nutrient regulation of signaling, transcription, and cell physiology by O-GlcNAcylation. , 2014, Cell metabolism.

[19]  Eun Jin Lee,et al.  Regulation of keratinocyte differentiation by O-GlcNAcylation. , 2014, Journal of dermatological science (Amsterdam).

[20]  Chi‐Huey Wong,et al.  O-GlcNAcylation regulates EZH2 protein stability and function , 2014, Proceedings of the National Academy of Sciences.

[21]  Sang-Cheol Lee,et al.  Src Family Kinase Inhibitor PP2 Has Different Effects on All-Trans-Retinoic Acid or Arsenic Trioxide-Induced Differentiation of an Acute Promyelocytic Leukemia Cell Line , 2013, Cancer research and treatment : official journal of Korean Cancer Association.

[22]  Ryan K. Dale,et al.  Ldb1-nucleated transcription complexes function as primary mediators of global erythroid gene activation. , 2013, Blood.

[23]  T. Bale,et al.  O-GlcNAc transferase (OGT) as a placental biomarker of maternal stress and reprogramming of CNS gene transcription in development , 2013, Proceedings of the National Academy of Sciences.

[24]  Christian J Stoeckert,et al.  Ontogeny of erythroid gene expression. , 2013, Blood.

[25]  E. Solary,et al.  TET2 and TET3 regulate GlcNAcylation and H3K4 methylation through OGT and SET1/COMPASS , 2013, The EMBO journal.

[26]  Xiaochun Yu,et al.  TET2 promotes histone O-GlcNAcylation during gene transcription , 2012, Nature.

[27]  Somasekar Seshagiri,et al.  Loss of the Tumor Suppressor BAP1 Causes Myeloid Transformation , 2012, Science.

[28]  G. Herrero-Beaumont,et al.  The Increase in O-Linked N-Acetylglucosamine Protein Modification Stimulates Chondrogenic Differentiation Both in Vitro and in Vivo* , 2012, The Journal of Biological Chemistry.

[29]  D. Rotin,et al.  LAPTM5 Protein Is a Positive Regulator of Proinflammatory Signaling Pathways in Macrophages* , 2012, The Journal of Biological Chemistry.

[30]  G. Hart,et al.  Evidence of the Involvement of O-GlcNAc-modified Human RNA Polymerase II CTD in Transcription in Vitro and in Vivo* , 2012, The Journal of Biological Chemistry.

[31]  Matthew S Macauley,et al.  Increasing O-GlcNAc slows neurodegeneration and stabilizes tau against aggregation. , 2012, Nature chemical biology.

[32]  R. Humphries,et al.  Functional Regulation of Pre-B-cell Leukemia Homeobox Interacting Protein 1 (PBXIP1/HPIP) in Erythroid Differentiation* , 2011, The Journal of Biological Chemistry.

[33]  Manolis Kellis,et al.  Dynamics of the epigenetic landscape during erythroid differentiation after GATA1 restoration. , 2011, Genome research.

[34]  L. Wood,et al.  Systemic upregulation of neutrophil α-defensins and serine proteases in neutrophilic asthma , 2011, Thorax.

[35]  R. Hardison,et al.  Bromodomain protein Brd3 associates with acetylated GATA1 to promote its chromatin occupancy at erythroid target genes , 2011, Proceedings of the National Academy of Sciences of the United States of America.

[36]  D. Tang,et al.  Autophagy regulates myeloid cell differentiation by p62/SQSTM1-mediated degradation of PML-RARα oncoprotein , 2011, Autophagy.

[37]  Edward L. Huttlin,et al.  A Tissue-Specific Atlas of Mouse Protein Phosphorylation and Expression , 2010, Cell.

[38]  G. Hart,et al.  β-N-acetylglucosamine (O-GlcNAc) is part of the histone code , 2010, Proceedings of the National Academy of Sciences.

[39]  Emery H. Bresnick,et al.  GATA Switches as Developmental Drivers* , 2010, The Journal of Biological Chemistry.

[40]  Christine Steinhoff,et al.  The genome-wide dynamics of the binding of Ldb1 complexes during erythroid differentiation. , 2010, Genes & development.

[41]  Francesca Chiaromonte,et al.  Erythroid GATA 1 function revealed by genome-wide analysis of transcription factor occupancy , histone modifications , and mRNA expression , 2009 .

[42]  Ernest Fraenkel,et al.  Insights into GATA-1-mediated gene activation versus repression via genome-wide chromatin occupancy analysis. , 2009, Molecular cell.

[43]  Henriette O'Geen,et al.  Discovering hematopoietic mechanisms through genome-wide analysis of GATA factor chromatin occupancy. , 2009, Molecular cell.

[44]  Kirby D. Johnson,et al.  Controlling hematopoiesis through sumoylation-dependent regulation of a GATA factor. , 2009, Molecular cell.

[45]  W. Moon,et al.  Excessive O‐GlcNAcylation of proteins suppresses spontaneous cardiogenesis in ES cells , 2009, FEBS letters.

[46]  R. Hardison,et al.  SCL and associated proteins distinguish active from repressive GATA transcription factor complexes. , 2008, Blood.

[47]  G. Davies,et al.  A potent mechanism-inspired O-GlcNAcase inhibitor that blocks phosphorylation of tau in vivo. , 2008, Nature chemical biology.

[48]  Steven P. Gygi,et al.  Large-scale phosphorylation analysis of mouse liver , 2007, Proceedings of the National Academy of Sciences.

[49]  G. Blobel,et al.  Acetylation of GATA-1 is required for chromatin occupancy. , 2006, Blood.

[50]  G. Blobel,et al.  Distinct Functions of Dispersed GATA Factor Complexes at an Endogenous Gene Locus , 2006, Molecular and Cellular Biology.

[51]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[52]  G. Hart,et al.  Perturbations in O-linked β-N-Acetylglucosamine Protein Modification Cause Severe Defects in Mitotic Progression and Cytokinesis* , 2005, Journal of Biological Chemistry.

[53]  Hao Wang,et al.  Global regulation of erythroid gene expression by transcription factor GATA-1. , 2004, Blood.

[54]  Paul T. Groth,et al.  The ENCODE (ENCyclopedia Of DNA Elements) Project , 2004, Science.

[55]  M. Daly,et al.  PGC-1α-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes , 2003, Nature Genetics.

[56]  Xiaoyong Yang,et al.  Recruitment of O-GlcNAc Transferase to Promoters by Corepressor mSin3A Coupling Protein O-GlcNAcylation to Transcriptional Repression , 2002, Cell.

[57]  S. Cameron,et al.  The zinc-finger proto-oncogene Gfi-1b is essential for development of the erythroid and megakaryocytic lineages. , 2002, Genes & development.

[58]  O. Nakajima,et al.  Heme deficiency in erythroid lineage causes differentiation arrest and cytoplasmic iron overload , 1999, The EMBO journal.

[59]  V. Ogryzko,et al.  Regulation of activity of the transcription factor GATA-1 by acetylation , 1998, Nature.

[60]  Y Fujiwara,et al.  Arrested development of embryonic red cell precursors in mouse embryos lacking transcription factor GATA-1. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[61]  J. Rowley,et al.  LAPTM5: a novel lysosomal-associated multispanning membrane protein preferentially expressed in hematopoietic cells. , 1996, Genomics.

[62]  M. Andreeff,et al.  Induction of differentiation in myeloid leukemia cell lines and acute promyelocytic leukemia cells by liposomal all-trans-retinoic acid. , 1993, Cancer research.

[63]  S. Orkin.  GATA-binding transcription factors in hematopoietic cells , 1992 .

[64]  S. Orkin,et al.  Erythroid differentiation in chimaeric mice blocked by a targeted mutation in the gene for transcription factor GATA-1 , 1991, Nature.

[65]  J. D. Engel,et al.  Activity and tissue-specific expression of the transcription factor NF-E1 multigene family. , 1990, Genes & development.

[66]  S. Wada,et al.  Terminal differentiation program of skeletal myogenesis is negatively regulated by O-GlcNAc glycosylation. , 2012, Biochimica et biophysica acta.

[67]  Yana Zhang,et al.  Core promoter sequence of SEMG I spans between the two putative GATA-1 binding domains and is responsive to IL-4 and IL-6 in myeloma cells. , 2009, Leukemia research.

[68]  K. Bomsztyk,et al.  Protocol for the fast chromatin immunoprecipitation (ChIP) method , 2006, Nature Protocols.

[69]  S. Orkin,et al.  GATA-binding transcription factors in hematopoietic cells. , 1992, Blood.