Noncoding RNA transcription targets AID to divergently transcribed loci in B cells

Nature. 2014 Oct 16;514(7522):389-93. doi: 10.1038/nature13580. Epub 2014 Aug 6.

Abstract

The vast majority of the mammalian genome has the potential to express noncoding RNA (ncRNA). The 11-subunit RNA exosome complex is the main source of cellular 3'-5' exoribonucleolytic activity and potentially regulates the mammalian noncoding transcriptome. Here we generated a mouse model in which the essential subunit Exosc3 of the RNA exosome complex can be conditionally deleted. Exosc3-deficient B cells lack the ability to undergo normal levels of class switch recombination and somatic hypermutation, two mutagenic DNA processes used to generate antibody diversity via the B-cell mutator protein activation-induced cytidine deaminase (AID). The transcriptome of Exosc3-deficient B cells has revealed the presence of many novel RNA exosome substrate ncRNAs. RNA exosome substrate RNAs include xTSS-RNAs, transcription start site (TSS)-associated antisense transcripts that can exceed 500 base pairs in length and are transcribed divergently from cognate coding gene transcripts. xTSS-RNAs are most strongly expressed at genes that accumulate AID-mediated somatic mutations and/or are frequent translocation partners of DNA double-strand breaks generated at Igh in B cells. Strikingly, translocations near TSSs or within gene bodies occur over regions of RNA exosome substrate ncRNA expression. These RNA exosome-regulated, antisense-transcribed regions of the B-cell genome recruit AID and accumulate single-strand DNA structures containing RNA-DNA hybrids. We propose that RNA exosome regulation of ncRNA recruits AID to single-strand DNA-forming sites of antisense and divergent transcription in the B-cell genome, thereby creating a link between ncRNA transcription and overall maintenance of B-cell genomic integrity.

Publication types

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

MeSH terms

  • Animals
  • B-Lymphocytes / metabolism*
  • Base Pairing
  • Cytidine Deaminase / metabolism*
  • DNA Breaks, Double-Stranded
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / metabolism
  • Exosome Multienzyme Ribonuclease Complex / deficiency
  • Exosome Multienzyme Ribonuclease Complex / genetics
  • Exosomes / metabolism
  • Female
  • Genome / genetics
  • Genomic Instability / genetics
  • Immunoglobulin Class Switching / genetics
  • Immunoglobulin Heavy Chains / genetics
  • Male
  • Mice
  • Nucleic Acid Hybridization
  • RNA, Antisense / biosynthesis
  • RNA, Antisense / chemistry
  • RNA, Antisense / genetics
  • RNA, Antisense / metabolism
  • RNA, Untranslated / biosynthesis*
  • RNA, Untranslated / chemistry
  • RNA, Untranslated / genetics*
  • RNA, Untranslated / metabolism
  • RNA-Binding Proteins / genetics
  • Somatic Hypermutation, Immunoglobulin / genetics
  • Substrate Specificity
  • Transcription Initiation Site
  • Transcription, Genetic / genetics*
  • Translocation, Genetic / genetics

Substances

  • DNA, Single-Stranded
  • Exosc3 protein, mouse
  • Immunoglobulin Heavy Chains
  • RNA, Antisense
  • RNA, Untranslated
  • RNA-Binding Proteins
  • Exosome Multienzyme Ribonuclease Complex
  • AICDA (activation-induced cytidine deaminase)
  • Cytidine Deaminase

Associated data

  • BioProject/PRJNA248775
  • SRA/SRP042355