title: Histone demethylase LSD1-mediated repression of GATA-2 is critical for erythroid differentiation.
abstract: BACKGROUND
  The transcription factor GATA-2 is predominantly expressed in hematopoietic stem and progenitor cells and counteracts the erythroid-specific transcription factor GATA-1, to modulate the proliferation and differentiation of hematopoietic cells. During hematopoietic cell differentiation, GATA-2 exhibits dynamic expression patterns, which are regulated by multiple transcription factors.
  
  
  METHODS
  Stable LSD1-knockdown cell lines were established by growing murine erythroleukemia (MEL) or mouse embryonic stem cells together with virus particles, in the presence of Polybrene(®) at 4 μg/mL, for 24-48 hours followed by puromycin selection (1 μg/mL) for 2 weeks. Real-time polymerase chain reaction (PCR)-based quantitative chromatin immunoprecipitation (ChIP) analysis was used to test whether the TAL1 transcription factor is bound to 1S promoter in the GATA-2 locus or whether LSD1 colocalizes with TAL1 at the 1S promoter. The sequential ChIP assay was utilized to confirm the role of LSD1 in the regulation of H3K4me2 at the GATA-2 locus during erythroid differentiation. Western blot analysis was employed to detect the protein expression. The alamarBlue(®) assay was used to examine the proliferation of the cells, and the absorbance was monitored at optical density (OD) 570 nm and OD 600 nm.
  
  
  RESULTS
  In this study, we showed that LSD1 regulates the expression of GATA-2 during erythroid differentiation. Knockdown of LSD1 results in increased GATA-2 expression and inhibits the differentiation of MEL and embryonic stem cells. Furthermore, we demonstrated that LSD1 binds to the 1S promoter of the GATA-2 locus and suppresses GATA-2 expression, via histone demethylation.
  
  
  CONCLUSIONS
  Our data revealed that LSD1 mediates erythroid differentiation, via epigenetic modification of the GATA-2 locus.
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Yidi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China ; Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun, People's Republic of China.
authors.forename: Xueqi
authors.initials: X
authors.lastname: Fu
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Jin
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Huo
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.affiliation: School of Life Sciences, Jilin University, Changchun, People's Republic of China ; Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun, People's Republic of China ; National Engineering Laboratory of AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Hu
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019175:DNA Methylation; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000201:enzymology*; D044127:Epigenesis, Genetic; D041905:Erythroid Cells / Q000201:enzymology*; D004920:Erythropoiesis; D050989:GATA2 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D056466:Histone Demethylases / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology* / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; D013997:Time Factors
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D050989:GATA2 Transcription Factor; D056466:Histone Demethylases; D006657:Histones; D011518:Proto-Oncogene Proteins; D000073942:T-Cell Acute Lymphocytic Leukemia Protein 1; C494712:Gata2 protein, mouse; C489366:Tal1 protein, mouse; C504076:KDM1a protein, mouse
keywords: GATA factor switching; LSD1; histone demethylation