title: ATRX, a member of the SNF2 family of helicase/ATPases, is required for chromosome alignment and meiotic spindle organization in metaphase II stage mouse oocytes.
abstract: ATRX is a centromeric heterochromatin binding protein belonging to the SNF2 family of helicase/ATPases with chromatin remodeling activity. Mutations in the human ATRX gene result in X-linked alpha-thalassaemia with mental retardation (ATRX) syndrome and correlate with changes in methylation of repetitive DNA sequences. We show here that ATRX also functions to regulate key stages of meiosis in mouse oocytes. At the germinal vesicle (GV) stage, ATRX was found associated with the perinucleolar heterochromatin rim in transcriptionally quiescent oocytes. Phosphorylation of ATRX during meiotic maturation is dependent upon calcium calmodulin kinase (CamKII) activity. Meiotic resumption also coincides with deacetylation of histone H4 at lysine 5 (H4K5 Ac) while ATRX and histone H3 methylated on lysine 9 (H3K9) remained bound to the centromeres and interstitial regions of condensing chromosomes, respectively. Inhibition of histone deacetylases (HDACs) with trichostatin A (TSA) disrupted ATRX binding to the centromeres of hyperacetylated chromosomes resulting in abnormal chromosome alignments at metaphase II (MII). Similarly, while selective ablation of ATRX by antibody microinjection and RNA interference (RNAi) had no effect on the progression of meiosis, it had severe consequences for the alignment of chromosomes on the metaphase II spindle. These results suggest that genome-wide epigenetic modifications such as global histone deacetylation are essential for the binding of ATRX to centromeric heterochromatin. Moreover, centromeric ATRX is required for correct chromosome alignment and organization of a bipolar meiotic metaphase II spindle.
authors.affiliation: The Jackson Laboratory, Bar Harbor, ME 04609, USA. rfuente@vet.upenn.edu
authors.forename: Rabindranath
authors.initials: R
authors.lastname: De La Fuente
authors.forename: Maria M
authors.initials: MM
authors.lastname: Viveiros
authors.forename: Karen
authors.initials: K
authors.lastname: Wigglesworth
authors.forename: John J
authors.initials: JJ
authors.lastname: Eppig
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002478:Cells, Cultured; D042002:Chromatin Assembly and Disassembly; D002875:Chromosomes / Q000235:genetics / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006570:Heterochromatin / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids / Q000494:pharmacology; D008239:Lysine / Q000378:metabolism; D008540:Meiosis / Q000502:physiology*; D008677:Metaphase / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D009865:Oocytes / Q000502:physiology*; D010766:Phosphorylation; D008941:Spindle Apparatus / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000075924:X-linked Nuclear Protein
chemical_list: D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D006570:Heterochromatin; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D009687:Nuclear Proteins; D014157:Transcription Factors; C012589:trichostatin A; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C513194:Smarca4 protein, mouse; D004265:DNA Helicases; C490462:Atrx protein, mouse; D000075924:X-linked Nuclear Protein; D008239:Lysine