Stabilization of ribozyme-like cis-noncoding rRNAs induces apoptotic and nonapoptotic death in lung cells

Cell Death Dis. 2012 Mar 15;3(3):e281. doi: 10.1038/cddis.2012.19.

Abstract

Bidirectional non-protein-coding RNAs are ubiquitously transcribed from the genome. Convergent sense and antisense transcripts may regulate each other. Here, we examined the convergent cis-noncoding rRNAs (nc-rRNAs) in A5 and E9 lung cancer models. Sense nc-rRNAs extending from rDNA intergenic region to internal transcribed spacer of around 10 kb in length were identified. nc-rRNAs in sense direction exhibited in vitro characteristics of ribozymes, namely, degradation upon incubation with MgCl(2) and stabilization by complementary oligonucleotides. Detection of endogenous cleavage-ligation products carrying internal deletion of hundreds to thousands nucleotides by massively parallel sequencing confirmed the catalytic properties. Transfection of oligonucleotides pairing with antisense nc-rRNAs stabilized both target and complementary transcripts, perturbed rRNA biogenesis, and induced massive cell death via apoptotic and/or nonapoptotic mechanisms depending on cell type and treatment. Oligonucleotides targeting cellular sense transcripts are less responsive. Spontaneously detached cells, though rare, also showed accumulation of nc-rRNAs and perturbation of rRNA biogenesis. Direct participation of nc-rRNAs in apoptotic and nonapoptotic death was demonstrated by transfection of synthetic nc-rRNAs encompassing the rDNA promoter. In sum, convergent cis-nc-rRNAs follow a feed-forward mechanism to regulate each other and rRNA biogenesis. This opens an opportunity to disrupt rRNA biogenesis, commonly upregulated in cancers, via inhibition of ribozyme-like activities in nc-rRNAs.

Publication types

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

MeSH terms

  • Adenocarcinoma
  • Adenocarcinoma of Lung
  • Animals
  • Apoptosis*
  • Base Sequence
  • Cell Cycle / genetics
  • Cell Death / genetics
  • Cell Line, Tumor
  • DNA, Ribosomal / genetics
  • DNA, Ribosomal / metabolism
  • Epithelial Cells / metabolism*
  • Epithelial Cells / pathology
  • High-Throughput Nucleotide Sequencing
  • Lung / metabolism*
  • Lung / pathology
  • Lung Neoplasms
  • Magnesium Chloride / metabolism
  • Mice
  • Molecular Sequence Data
  • Oligonucleotides, Antisense / genetics
  • Oligonucleotides, Antisense / metabolism
  • Promoter Regions, Genetic
  • RNA, Catalytic / genetics
  • RNA, Catalytic / metabolism*
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism*
  • RNA, Untranslated / genetics
  • RNA, Untranslated / metabolism*
  • Signal Transduction
  • Transcription, Genetic
  • Transfection

Substances

  • DNA, Ribosomal
  • Oligonucleotides, Antisense
  • RNA, Catalytic
  • RNA, Ribosomal
  • RNA, Untranslated
  • Magnesium Chloride