title: Role of ALKBH1 in the Core Transcriptional Network of Embryonic Stem Cells.
abstract: OBJECTIVE
  ALKBH1, an AlkB homologue in the 2-oxoglutarate and Fe2+ dependent hydroxylase family, is a histone dioxygenase that removes methyl groups from histone H2A. Studies of transgenic mice lacking Alkbh1 reveal that most Alkbh1-/- embryos die during embryonic development. Embryonic stem cells (ESCs) derived from these mice have prolonged expression of pluripotency markers and delayed induction of genes involved in neural differentiation, indicating that ALKBH1 is involved in regulation of pluripotency and differentiation. The aim of this study was to further investigate the role ALKBH1 in early development.
  
  
  METHODS
  Double-filter methods for nitrocellulose-filter binding, dot blot, enzyme-linked immunosorbent assay (ELISA), immonocytochemistry, cell culture and differentiation of mouse ESCs, Co-IP and miRNA analysis.
  
  
  RESULTS
  We found that SOX2 and NANOG bind the ALKBH1 promoter, and we identified protein-protein interactions between ALKBH1 and these core transcription factors of the pluripotency network. Furthermore, lack of ALKBH1 affected the expression of developmentally important miRNAs, which are involved in the regulation of NANOG, SOX2 and neural differentiation.
  
  
  CONCLUSIONS
  Our results suggest that ALKBH1 interacts with the core transcriptional pluripotency network of ESCs and is involved in regulation of pluripotency and differentiation.
authors.affiliation: Clinic for Diagnostics and Intervention and Institute of Medical Microbiology, Oslo University Hospital, Rikshospitalet, Oslo, Norway.
authors.forename: Rune
authors.initials: R
authors.lastname: Ougland
authors.forename: Ida
authors.initials: I
authors.lastname: Jonson
authors.forename: Marivi Nabong
authors.initials: MN
authors.lastname: Moen
authors.forename: Gaute
authors.initials: G
authors.lastname: Nesse
authors.forename: Gry
authors.initials: G
authors.lastname: Asker
authors.forename: Arne
authors.initials: A
authors.lastname: Klungland
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Larsen
mesh_terms: D020121:5' Untranslated Regions; D000071498:AlkB Homolog 1, Histone H2a Dioxygenase; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002454:Cell Differentiation; D002460:Cell Line; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D055786:Gene Knockout Techniques; D053263:Gene Regulatory Networks; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000737:chemistry / Q000378:metabolism; D011401:Promoter Regions, Genetic; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D020121:5' Untranslated Regions; D018398:Homeodomain Proteins; D035683:MicroRNAs; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011994:Recombinant Proteins; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D014157:Transcription Factors; D000071498:AlkB Homolog 1, Histone H2a Dioxygenase; C529511:Alkbh1 protein, mouse; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase