Loss of full-length hnRNP R isoform impairs DNA damage response in motoneurons by inhibiting Yb1 recruitment to chromatin

Nucleic Acids Res. 2021 Dec 2;49(21):12284-12305. doi: 10.1093/nar/gkab1120.

Abstract

Neurons critically rely on the functions of RNA-binding proteins to maintain their polarity and resistance to neurotoxic stress. HnRNP R has a diverse range of post-transcriptional regulatory functions and is important for neuronal development by regulating axon growth. Hnrnpr pre-mRNA undergoes alternative splicing giving rise to a full-length protein and a shorter isoform lacking its N-terminal acidic domain. To investigate functions selectively associated with the full-length hnRNP R isoform, we generated a Hnrnpr knockout mouse (Hnrnprtm1a/tm1a) in which expression of full-length hnRNP R was abolished while production of the truncated hnRNP R isoform was retained. Motoneurons cultured from Hnrnprtm1a/tm1a mice did not show any axonal growth defects but exhibited enhanced accumulation of double-strand breaks and an impaired DNA damage response upon exposure to genotoxic agents. Proteomic analysis of the hnRNP R interactome revealed the multifunctional protein Yb1 as a top interactor. Yb1-depleted motoneurons were defective in DNA damage repair. We show that Yb1 is recruited to chromatin upon DNA damage where it interacts with γ-H2AX, a mechanism that is dependent on full-length hnRNP R. Our findings thus suggest a novel role of hnRNP R in maintaining genomic integrity and highlight the function of its N-terminal acidic domain in this context.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / metabolism
  • Cell Line
  • Cells, Cultured
  • Chromatin / genetics*
  • Chromatin / metabolism
  • DNA Damage*
  • DNA Repair / genetics*
  • HEK293 Cells
  • Heterogeneous-Nuclear Ribonucleoproteins / genetics*
  • Heterogeneous-Nuclear Ribonucleoproteins / metabolism
  • Humans
  • Immunoblotting
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Motor Neurons / cytology
  • Motor Neurons / metabolism*
  • Protein Binding
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Y-Box-Binding Protein 1 / genetics*
  • Y-Box-Binding Protein 1 / metabolism

Substances

  • Chromatin
  • Heterogeneous-Nuclear Ribonucleoproteins
  • Hnrpr protein, mouse
  • Protein Isoforms
  • Y-Box-Binding Protein 1