title: Alternative mechanisms associated with silencing of CDKN1C in Beckwith-Wiedemann syndrome.
abstract: BACKGROUND
  Mutations in the imprinted gene CDKN1C account for approximately 10% of Beckwith-Wiedemann syndrome (BWS) cases. Fibroblasts from BWS patients with loss of methylation (LOM) at the imprinting control region (ICR) KvDMR1 have reduced CDKN1C expression. Another group of BWS patients with downregulated CDKN1C expression but with normal methylation at KvDMR1 has been identified.
  
  
  OBJECTIVE
  To investigate the mechanism of CDKN1C silencing in BWS in these two classes of patients.
  
  
  METHODS
  The CDKN1C promoter region was analysed for changes in DNA methylation using bisulphite sequencing, and for alterations in chromatin structure using the chromatin immunoprecipitation (ChIP) assay.
  
  
  RESULTS
  There was only spurious CpG methylation of the CDKN1C promoter in fibroblast DNA from both normal individuals and patients with BWS, irrespective of the methylation status of KvDMR1. There was no detectable change in chromatin structure at the CDKN1C promoter in patients with LOM at KvDMR1. BWS patients with downregulated CDKN1C and normal methylation at KvDMR1 had depletion of dimethylated H3-K4 and enrichment of dimethylated H3-K9 and HP1gamma at the CDKN1C promoter, suggesting that in these cases gene silencing is associated with repressive chromatin changes.
  
  
  CONCLUSIONS
  CDKN1C may be downregulated by multiple mechanisms including some that do not involve promoter methylation. In BWS patients with normal methylation at KvDMR1 and reduced expression of CDKN1C, repressive chromatin may play a role, but the absence of methylation and repressive chromatin structure at the CDKN1C promoter in BWS patients with LOM at KvDMR1 argues for a direct role of this epimutation in silencing CDKN1C.
authors.affiliation: Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
authors.forename: N
authors.initials: N
authors.lastname: Diaz-Meyer
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: S N
authors.initials: SN
authors.lastname: Sait
authors.forename: E R
authors.initials: ER
authors.lastname: Maher
authors.forename: M J
authors.initials: MJ
authors.lastname: Higgins
mesh_terms: D001506:Beckwith-Wiedemann Syndrome / Q000235:genetics*; D002843:Chromatin / Q000648:ultrastructure; D050761:Cyclin-Dependent Kinase Inhibitor p57 / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation; D015536:Down-Regulation; D020868:Gene Silencing; D018392:Genomic Imprinting; D006801:Humans; D011401:Promoter Regions, Genetic
chemical_list: C493964:CDKN1C protein, human; D002843:Chromatin; D050761:Cyclin-Dependent Kinase Inhibitor p57