Identifying Centromeric RNAs Involved in Histone Dynamics In Vivo

Methods Enzymol. 2016:573:445-66. doi: 10.1016/bs.mie.2016.01.010. Epub 2016 Mar 24.

Abstract

Over the last decade, the long accepted dogma that heterochromatin is silent has been challenged by increasing evidence of active transcription in these apocryphally annotated quiescent regions of the genome. The recent discovery of noncoding RNAs (ncRNAs) originating from, or localizing to, centromeres, pericentromeres, and telomeres (ie, constitutive heterochromatin) suggest a potential role for ncRNAs in genome integrity. This new paradigm suggests that ncRNAs may recruit chromatin-binding factors, stabilize the higher order folded state of the chromatin fiber, and participate in regulation of processes such as transcription-mediated nucleosome assembly. Thus, identifying, purifying, and elucidating the function of ncRNAs has the potential to provide key insights into genome organization and is currently a topic of intense experimental investigation.

Keywords: CENP-A; Centromere; Genome organization; Histone variants; Noncoding RNA; RNA FISH; RNA-Seq.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Autoantigens / analysis
  • Autoantigens / metabolism
  • Cell Culture Techniques / methods
  • Centromere / chemistry
  • Centromere / metabolism*
  • Centromere Protein A
  • Chromatin / chemistry
  • Chromatin / metabolism
  • Chromosomal Proteins, Non-Histone / analysis
  • Chromosomal Proteins, Non-Histone / metabolism
  • Fluorescent Antibody Technique / methods
  • HeLa Cells
  • Histones / analysis
  • Histones / metabolism*
  • Humans
  • Microscopy, Fluorescence / methods
  • RNA, Untranslated / analysis
  • RNA, Untranslated / genetics
  • RNA, Untranslated / metabolism*
  • Sequence Analysis, RNA / methods

Substances

  • Autoantigens
  • CENPA protein, human
  • Centromere Protein A
  • Chromatin
  • Chromosomal Proteins, Non-Histone
  • Histones
  • RNA, Untranslated