MicroRNAs down-regulate homologous recombination in the G1 phase of cycling cells to maintain genomic stability

Elife. 2014 Apr 30:3:e02445. doi: 10.7554/eLife.02445.

Abstract

Homologous recombination (HR)-mediated repair of DNA double-strand break (DSB)s is restricted to the post-replicative phases of the cell cycle. Initiation of HR in the G1 phase blocks non-homologous end joining (NHEJ) impairing DSB repair. Completion of HR in G1 cells can lead to the loss-of-heterozygosity (LOH), which is potentially carcinogenic. We conducted a gain-of-function screen to identify miRNAs that regulate HR-mediated DSB repair, and of these miRNAs, miR-1255b, miR-148b*, and miR-193b* specifically suppress the HR-pathway in the G1 phase. These miRNAs target the transcripts of HR factors, BRCA1, BRCA2, and RAD51, and inhibiting miR-1255b, miR-148b*, and miR-193b* increases expression of BRCA1/BRCA2/RAD51 specifically in the G1-phase leading to impaired DSB repair. Depletion of CtIP, a BRCA1-associated DNA end resection protein, rescues this phenotype. Furthermore, deletion of miR-1255b, miR-148b*, and miR-193b* in independent cohorts of ovarian tumors correlates with significant increase in LOH events/chromosomal aberrations and BRCA1 expression.DOI: http://dx.doi.org/10.7554/eLife.02445.001.

Keywords: BRCA1; DNA repair; cell cycle.

Publication types

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

MeSH terms

  • Base Sequence
  • Cell Line, Tumor
  • DNA Breaks, Double-Stranded
  • Down-Regulation / drug effects
  • Down-Regulation / genetics*
  • Enzyme Inhibitors / pharmacology
  • Female
  • G1 Phase / drug effects
  • G1 Phase / genetics*
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Library
  • Genomic Instability*
  • Homologous Recombination / drug effects
  • Homologous Recombination / genetics*
  • Humans
  • Loss of Heterozygosity / genetics
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Models, Genetic
  • Molecular Sequence Data
  • Ovarian Neoplasms / genetics
  • Ovarian Neoplasms / pathology
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Poly(ADP-ribose) Polymerases / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Recombinational DNA Repair
  • Triple Negative Breast Neoplasms / genetics
  • Triple Negative Breast Neoplasms / pathology

Substances

  • Enzyme Inhibitors
  • MicroRNAs
  • Poly(ADP-ribose) Polymerase Inhibitors
  • RNA, Messenger
  • Poly(ADP-ribose) Polymerases