Human fetal globin gene expression is regulated by LYAR

Human globin gene expression during development is modulated by transcription factors in a stage-dependent manner. However, the mechanisms controlling the process are still largely unknown. In this study, we found that a nuclear protein, LYAR (human homologue of mouse Ly-1 antibody reactive clone) directly interacted with the methyltransferase PRMT5 which triggers the histone H4 Arg3 symmetric dimethylation (H4R3me2s) mark. We found that PRMT5 binding on the proximal γ-promoter was LYAR-dependent. The LYAR DNA-binding motif (GGTTAT) was identified by performing CASTing (cyclic amplification and selection of targets) experiments. Results of EMSA and ChIP assays confirmed that LYAR bound to a DNA region corresponding to the 5′-untranslated region of the γ-globin gene. We also found that LYAR repressed human fetal globin gene expression in both K562 cells and primary human adult erythroid progenitor cells. Thus, these data indicate that LYAR acts as a novel transcription factor that binds the γ-globin gene, and is essential for silencing the γ-globin gene.

[1]  S. Orkin,et al.  Transcriptional silencing of {gamma}-globin by BCL11A involves long-range interactions and cooperation with SOX6. , 2010, Genes & development.

[2]  Aiping Zhang,et al.  Crystal structure of the human PRMT5:MEP50 complex , 2012, Proceedings of the National Academy of Sciences.

[3]  W. Funk,et al.  CASTing for multicomponent DNA-binding complexes. , 1993, Trends in biochemical sciences.

[4]  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.

[5]  Daniel E Bauer,et al.  Reawakening fetal hemoglobin: prospects for new therapies for the β-globin disorders. , 2012, Blood.

[6]  S. Jane,et al.  Induction of Human Fetal Globin Gene Expression by a Novel Erythroid Factor, NF-E4 , 2000, Molecular and Cellular Biology.

[7]  S. Fucharoen,et al.  A novel C-T transition within the distal CCAAT motif of the G gamma-globin gene in the Japanese HPFH: implication of factor binding in elevated fetal globin expression. , 1990, Nucleic acids research.

[8]  G. Stamatoyannopoulos,et al.  FKLF, a Novel Krüppel-Like Factor That Activates Human Embryonic and Fetal β-Like Globin Genes , 1999, Molecular and Cellular Biology.

[9]  G. Stamatoyannopoulos,et al.  FKLF-2: a novel Krüppel-like transcriptional factor that activates globin and other erythroid lineage genes. , 2000, Blood.

[10]  G. D. Ginder,et al.  Methyl binding domain protein 2 mediates gamma-globin gene silencing in adult human betaYAC transgenic mice. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[11]  B. Pace,et al.  Understanding mechanisms of γ‐globin gene regulation to develop strategies for pharmacological fetal hemoglobin induction , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[12]  B. Pace,et al.  Globin Gene Silencing in Primary Erythroid Cultures , 1997, The Journal of Biological Chemistry.

[13]  S. Jane,et al.  Hemoglobin switching in man and chicken is mediated by a heteromeric complex between the ubiquitous transcription factor CP2 and a developmentally specific protein. , 1995, The EMBO journal.

[14]  G. Hong,et al.  Nucleic Acids Research , 2015, Nucleic Acids Research.

[15]  M. Trudel,et al.  Ikaros and GATA-1 Combinatorial Effect Is Required for Silencing of Human γ-Globin Genes , 2008, Molecular and Cellular Biology.

[16]  A. Cantor,et al.  Role of ZBP-89 in human globin gene regulation and erythroid differentiation. , 2011, Blood.

[17]  K. Gaensler,et al.  Methyl binding domain protein 2 mediates γ-globin gene silencing in adult human βYAC transgenic mice , 2006 .

[18]  Jiahai Zhang,et al.  Solution structure of an atypical PHD finger in BRPF2 and its interaction with DNA. , 2012, Journal of structural biology.

[19]  F. Grosveld,et al.  Defective haematopoiesis in fetal liver resulting from inactivation of the EKLF gene , 1995, Nature.

[20]  Li Liu,et al.  Role of STAT3 and GATA-1 interactions in gamma-globin gene expression. , 2009, Experimental Hematology.

[21]  K. Betke,et al.  Estimation of small percentages of foetal haemoglobin. , 1959, Nature.

[22]  J. Strouboulis,et al.  Erythropoiesis: Model systems, molecular regulators, and developmental programs , 2009, IUBMB life.

[23]  J. Bieker,et al.  Analyses of beta-thalassemia mutant DNA interactions with erythroid Krüppel-like factor (EKLF), an erythroid cell-specific transcription factor. , 1994, The Journal of biological chemistry.

[24]  S. Orkin,et al.  Lethal β-thalassaemia in mice lacking the erythroid CACCC-transcription factor EKLF , 1995, Nature.

[25]  F. Grosveld,et al.  Fetal globin expression is regulated by Friend of Prmt1. , 2010, Blood.

[26]  J. D. Engel,et al.  An embryonic/fetal β‐type globin gene repressor contains a nuclear receptor TR2/TR4 heterodimer , 2002, The EMBO journal.

[27]  K. Betke,et al.  Estimation of Small Percentages of Fœtal Hæmoglobin , 1959, Nature.

[28]  T. Mercer,et al.  Transcriptional effects of a lupus-associated polymorphism in the 5' untranslated region (UTR) of human complement receptor 2 (CR2/CD21). , 2012, Molecular immunology.

[29]  John Loughlin,et al.  An SNP in the 5'-UTR of GDF5 is associated with osteoarthritis susceptibility in Europeans and with in vivo differences in allelic expression in articular cartilage. , 2007, Human molecular genetics.

[30]  M. Steinberg,et al.  BCL11A represses HBG transcription in K562 cells. , 2009, Blood cells, molecules & diseases.

[31]  D. Tuan,et al.  NF-Y Recruits Both Transcription Activator and Repressor to Modulate Tissue- and Developmental Stage-Specific Expression of Human γ-Globin Gene , 2012, PloS one.

[32]  J. Hirschhorn,et al.  Supporting Online Material Materials and Methods Figs. S1 to S10 Tables S1 to S7 References Human Fetal Hemoglobin Expression Is Regulated by the Developmental Stage-specific Repressor Bcl11a , 2022 .

[33]  G. Stamatoyannopoulos.  Control of globin gene expression during development and erythroid differentiation. , 2005, Experimental hematology.

[34]  Robert L Moritz,et al.  PRMT5-mediated methylation of histone H4R3 recruits DNMT3A, coupling histone and DNA methylation in gene silencing , 2009, Nature Structural &Molecular Biology.

[35]  J. Bieker.  Probing the onset and regulation of erythroid cell-specific gene expression. , 2005, The Mount Sinai journal of medicine, New York.

[36]  N. Olivieri,et al.  Two novel beta-thalassemia mutations in the 5' and 3' noncoding regions of the beta-globin gene. , 1992, Blood.

[37]  E. Dong,et al.  Analysis of the GAD1 promoter: Trans-acting factors and DNA methylation converge on the 5′ untranslated region , 2011, Neuropharmacology.

[38]  I. Weissman,et al.  LYAR, a novel nucleolar protein with zinc finger DNA-binding motifs, is involved in cell growth regulation. , 1993, Genes & development.

[39]  H. Hammad,et al.  From bloodjournal.hematologylibrary.org at PENN STATE UNIVERSITY on February 22, 2013. For personal use only. , 2005 .

[40]  D. Weatherall.  Thalassemia as a global health problem: recent progress toward its control in the developing countries , 2010, Annals of the New York Academy of Sciences.

[41]  A. Bank.  Regulation of human fetal hemoglobin: new players, new complexities. , 2006, Blood.

[42]  Ming-Ming Zhou,et al.  The PHD finger: a versatile epigenome reader. , 2011, Trends in biochemical sciences.

[43]  Steve Best,et al.  cMYB is involved in the regulation of fetal hemoglobin production in adults. , 2006, Blood.

[44]  J. D. Engel,et al.  Forced TR2/TR4 expression in sickle cell disease mice confers enhanced fetal hemoglobin synthesis and alleviated disease phenotypes , 2011, Proceedings of the National Academy of Sciences.

[45]  Yingming Zhao,et al.  Ly‐1 Antibody Reactive Clone Is an Important Nucleolar Protein for Control of Self‐Renewal and Differentiation in Embryonic Stem Cells , 2009, Stem cells.

[46]  J. Hoyer,et al.  ICSH recommendations for the measurement of Haemoglobin F , 2012, International journal of laboratory hematology.

[47]  Stuart H. Orkin,et al.  Developmental and species-divergent globin switching are driven by BCL11A , 2009, Nature.

[48]  J. Hoyer,et al.  ICSH recommendations for the measurement of Haemoglobin A2 , 2012, International journal of laboratory hematology.

[49]  Xingxu Huang,et al.  Synergistic Effect of SRY and Its Direct Target, WDR5, on Sox9 Expression , 2012, PloS one.

[50]  Mark A. Hall,et al.  The Role of p22 NF-E4 in Human Globin Gene Switching* , 2004, Journal of Biological Chemistry.

[51]  K. Zhao,et al.  Attenuation of Forkhead signaling by the retinal determination factor DACH1 , 2010, Proceedings of the National Academy of Sciences.

[52]  R. Moritz,et al.  Identification of a PRMT5-dependent repressor complex linked to silencing of human fetal globin gene expression. , 2010, Blood.

[53]  B. A. Lewis,et al.  A downstream element in the human beta-globin promoter: evidence of extended sequence-specific transcription factor IID contacts. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[54]  W. Pierce,et al.  AS1411 alters the localization of a complex containing protein arginine methyltransferase 5 and nucleolin. , 2007, Cancer research.

[55]  Masayuki Yamamoto,et al.  GATA1 Function, a Paradigm for Transcription Factors in Hematopoiesis , 2005, Molecular and Cellular Biology.