Escape From X Inactivation Varies in Mouse Tissues

PLoS Genet. 2015 Mar 18;11(3):e1005079. doi: 10.1371/journal.pgen.1005079. eCollection 2015 Mar.

Abstract

X chromosome inactivation (XCI) silences most genes on one X chromosome in female mammals, but some genes escape XCI. To identify escape genes in vivo and to explore molecular mechanisms that regulate this process we analyzed the allele-specific expression and chromatin structure of X-linked genes in mouse tissues and cells with skewed XCI and distinguishable alleles based on single nucleotide polymorphisms. Using a binomial model to assess allelic expression, we demonstrate a continuum between complete silencing and expression from the inactive X (Xi). The validity of the RNA-seq approach was verified using RT-PCR with species-specific primers or Sanger sequencing. Both common escape genes and genes with significant differences in XCI status between tissues were identified. Such genes may be candidates for tissue-specific sex differences. Overall, few genes (3-7%) escape XCI in any of the mouse tissues examined, suggesting stringent silencing and escape controls. In contrast, an in vitro system represented by the embryonic-kidney-derived Patski cell line showed a higher density of escape genes (21%), representing both kidney-specific escape genes and cell-line specific escape genes. Allele-specific RNA polymerase II occupancy and DNase I hypersensitivity at the promoter of genes on the Xi correlated well with levels of escape, consistent with an open chromatin structure at escape genes. Allele-specific CTCF binding on the Xi clustered at escape genes and was denser in brain compared to the Patski cell line, possibly contributing to a more compartmentalized structure of the Xi and fewer escape genes in brain compared to the cell line where larger domains of escape were observed.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CCCTC-Binding Factor
  • Deoxyribonuclease I / metabolism
  • Female
  • Mice
  • Organ Specificity
  • Polymorphism, Single Nucleotide
  • RNA Polymerase II / metabolism
  • Repressor Proteins / metabolism
  • Sequence Analysis, RNA
  • X Chromosome Inactivation*

Substances

  • CCCTC-Binding Factor
  • Ctcf protein, mouse
  • Repressor Proteins
  • RNA Polymerase II
  • Deoxyribonuclease I