PolyC-binding proteins enhance expression of the CDK2 cell cycle regulatory protein via alternative splicing

Nucleic Acids Res. 2018 Feb 28;46(4):2030-2044. doi: 10.1093/nar/gkx1255.

Abstract

The PolyC binding proteins (PCBPs) impact alternative splicing of a subset of mammalian genes that are enriched in basic cellular functions. Here, we focus our analysis on PCBP-controlled cassette exon-splicing within the cell cycle control regulator cyclin-dependent kinase-2 (CDK2) transcript. We demonstrate that PCBP binding to a C-rich polypyrimidine tract (PPT) preceding exon 5 of the CDK2 transcript enhances cassette exon inclusion. This splice enhancement is U2AF65-independent and predominantly reflects actions of the PCBP1 isoform. Remarkably, PCBPs' control of CDK2 ex5 splicing has evolved subsequent to mammalian divergence via conversion of constitutive exon 5 inclusion in the mouse CDK2 transcript to PCBP-responsive exon 5 alternative splicing in humans. Importantly, exclusion of exon 5 from the hCDK2 transcript dramatically represses the expression of CDK2 protein with a corresponding perturbation in cell cycle kinetics. These data highlight a recently evolved post-transcriptional pathway in primate species with the potential to modulate cell cycle control.

Publication types

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

MeSH terms

  • Alternative Splicing*
  • Animals
  • Cell Cycle
  • Cyclin-Dependent Kinase 2 / genetics*
  • Cyclin-Dependent Kinase 2 / metabolism
  • DNA-Binding Proteins
  • Evolution, Molecular
  • Exons
  • Heterogeneous-Nuclear Ribonucleoproteins / metabolism
  • Humans
  • K562 Cells
  • Mice
  • Polypyrimidine Tract-Binding Protein / metabolism
  • RNA Splicing Factors / metabolism*
  • RNA-Binding Proteins
  • Splicing Factor U2AF / metabolism

Substances

  • DNA-Binding Proteins
  • Heterogeneous-Nuclear Ribonucleoproteins
  • PCBP1 protein, human
  • RNA Splicing Factors
  • RNA-Binding Proteins
  • Splicing Factor U2AF
  • U2AF2 protein, human
  • Polypyrimidine Tract-Binding Protein
  • CDK2 protein, human
  • Cdk2 protein, mouse
  • Cyclin-Dependent Kinase 2