Coilin displays differential affinity for specific RNAs in vivo and is linked to telomerase RNA biogenesis

J Mol Biol. 2013 Feb 22;425(4):713-24. doi: 10.1016/j.jmb.2012.12.014. Epub 2012 Dec 27.

Abstract

Coilin is widely known as the protein marker of the Cajal body, a subnuclear domain important to the biogenesis of small nuclear ribonucleoproteins and telomerase, complexes that are crucial to pre-messenger RNA splicing and telomere maintenance, respectively. Extensive studies have characterized the interaction between coilin and the various other protein components of CBs and related subnuclear domains; however, only a few have examined interactions between coilin and nucleic acid. We have recently published that coilin is tightly associated with nucleic acid, displays RNase activity in vitro, and is redistributed to the ribosomal RNA (rRNA)-rich nucleoli in cells treated with the DNA-damaging agents cisplatin and etoposide. Here, we report a specific in vivo association between coilin and rRNA, U small nuclear RNA (snRNA), and human telomerase RNA, which is altered upon treatment with DNA-damaging agents. Using chromatin immunoprecipitation, we provide evidence of coilin interaction with specific regions of U snRNA gene loci. We have also utilized bacterially expressed coilin fragments in order to map the region(s) important for RNA binding and RNase activity in vitro. Additionally, we provide evidence of coilin involvement in the processing of human telomerase RNA both in vitro and in vivo.

Publication types

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

MeSH terms

  • Binding Sites
  • Binding, Competitive
  • Cell Line
  • Chromatin Immunoprecipitation
  • Coiled Bodies / metabolism
  • DNA / genetics
  • DNA / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • HeLa Cells
  • Humans
  • Mutation
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Protein Binding
  • RNA / genetics
  • RNA / metabolism*
  • RNA Interference
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism*
  • RNA, Small Nuclear / genetics
  • RNA, Small Nuclear / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ribonucleases / genetics
  • Ribonucleases / metabolism
  • Telomerase / genetics

Substances

  • Nuclear Proteins
  • RNA, Ribosomal
  • RNA, Small Nuclear
  • p80-coilin
  • RNA
  • DNA
  • Telomerase
  • Ribonucleases