HUWE1 interacts with PCNA to alleviate replication stress

EMBO Rep. 2016 Jun;17(6):874-86. doi: 10.15252/embr.201541685. Epub 2016 May 4.

Abstract

Defects in DNA replication, DNA damage response, and DNA repair compromise genomic stability and promote cancer development. In particular, unrepaired DNA lesions can arrest the progression of the DNA replication machinery during S-phase, causing replication stress, mutations, and DNA breaks. HUWE1 is a HECT-type ubiquitin ligase that targets proteins involved in cell fate, survival, and differentiation. Here, we report that HUWE1 is essential for genomic stability, by promoting replication of damaged DNA We show that HUWE1-knockout cells are unable to mitigate replication stress, resulting in replication defects and DNA breakage. Importantly, we find that this novel role of HUWE1 requires its interaction with the replication factor PCNA, a master regulator of replication fork restart, at stalled replication forks. Finally, we provide evidence that HUWE1 mono-ubiquitinates H2AX to promote signaling at stalled forks. Altogether, our work identifies HUWE1 as a novel regulator of the replication stress response.

Keywords: DNA replication; H2AX; HUWE1; PCNA; genomic instability.

MeSH terms

  • Cell Line
  • DNA Damage
  • DNA Repair
  • DNA Replication*
  • Gene Knockout Techniques
  • Genomic Instability
  • Histones / metabolism
  • Humans
  • Phenotype
  • Proliferating Cell Nuclear Antigen / metabolism*
  • Protein Binding
  • Protein Processing, Post-Translational
  • Protein Transport
  • Stress, Physiological*
  • Tumor Suppressor Proteins
  • Ubiquitin / metabolism
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination

Substances

  • Histones
  • Proliferating Cell Nuclear Antigen
  • Tumor Suppressor Proteins
  • Ubiquitin
  • HUWE1 protein, human
  • Ubiquitin-Protein Ligases