Nuclear PKM2 binds pre-mRNA at folded G-quadruplexes and reveals their gene regulatory role

Mol Cell. 2024 Oct 3;84(19):3775-3789.e6. doi: 10.1016/j.molcel.2024.07.025. Epub 2024 Aug 16.

Abstract

Nuclear localization of the metabolic enzyme PKM2 is widely observed in various cancer types. We identify nuclear PKM2 as a non-canonical RNA-binding protein (RBP) that specifically interacts with folded RNA G-quadruplex (rG4) structures in precursor mRNAs (pre-mRNAs). PKM2 occupancy at rG4s prevents the binding of repressive RBPs, such as HNRNPF, and promotes the expression of rG4-containing pre-mRNAs (the "rG4ome"). We observe an upregulation of the rG4ome during epithelial-to-mesenchymal transition and a negative correlation of rG4 abundance with patient survival in different cancer types. By preventing the nuclear accumulation of PKM2, we could repress the rG4ome in triple-negative breast cancer cells and reduce migration and invasion of cancer cells in vitro and in xenograft mouse models. Our data suggest that the balance of folded and unfolded rG4s controlled by RBPs impacts gene expression during tumor progression.

Keywords: G-quadruplex; G4; PAR-CLIP; PKM2; RNA-binding proteins; crosslinking and immunoprecipitation; posttranscriptional gene regulation.

MeSH terms

  • Animals
  • Carrier Proteins* / genetics
  • Carrier Proteins* / metabolism
  • Cell Line, Tumor
  • Cell Movement
  • Cell Nucleus* / genetics
  • Cell Nucleus* / metabolism
  • Epithelial-Mesenchymal Transition* / genetics
  • Female
  • G-Quadruplexes*
  • Gene Expression Regulation, Neoplastic*
  • HEK293 Cells
  • Heterogeneous-Nuclear Ribonucleoprotein Group F-H / genetics
  • Heterogeneous-Nuclear Ribonucleoprotein Group F-H / metabolism
  • Humans
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Mice
  • Mice, Inbred NOD
  • Neoplasm Invasiveness
  • Protein Binding
  • RNA Precursors* / genetics
  • RNA Precursors* / metabolism
  • Thyroid Hormone-Binding Proteins*
  • Thyroid Hormones* / genetics
  • Thyroid Hormones* / metabolism
  • Triple Negative Breast Neoplasms / genetics
  • Triple Negative Breast Neoplasms / metabolism
  • Triple Negative Breast Neoplasms / pathology

Substances

  • Carrier Proteins
  • Heterogeneous-Nuclear Ribonucleoprotein Group F-H
  • Membrane Proteins
  • RNA Precursors
  • Thyroid Hormone-Binding Proteins
  • Thyroid Hormones
  • PKM2 protein, human