NF-YA enters cells through cell penetrating peptides

Biochim Biophys Acta Mol Cell Res. 2019 Mar;1866(3):430-440. doi: 10.1016/j.bbamcr.2018.10.004. Epub 2018 Oct 6.

Abstract

Cell Penetrating Peptides -CPPs- are short aminoacidic stretches present in proteins that have the ability to translocate the plasma membrane and facilitate delivery of various molecules. They are usually rich in basic residues, and organized as alpha helices. NF-Y is a transcription factor heterotrimer formed by two Histone Fold Domain -HFD- subunits and the sequence-specific NF-YA. NF-YA possesses two α-helices rich in basic residues. We show that it efficiently enters cells at nanomolar concentrations in the absence of carrier peptides. Mutagenesis identified at least two separate CPPs in the A1 and A2, which overlap with previously identified nuclear localization signals (NLS). The half-life of the transduced protein is short in human cancer cells, longer in mouse C2C12 myoblasts. The internalized NF-YA is capable of trimerization with the HFD subunits and binding to the target CCAAT box. Functionality is further suggested by protein transfection in C2C12 cells, leading to inhibition of differentiation to myotubes. In conclusion, NF-YA contains CPPs, hinting at novel -and unexpected- properties of this subunit.

Keywords: CCAAT; Cell penetrating peptide; NF-Y; Transcription factor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CCAAT-Binding Factor / genetics
  • CCAAT-Binding Factor / metabolism*
  • Cell Line
  • Cell Line, Tumor
  • Cell-Penetrating Peptides / metabolism*
  • DNA-Binding Proteins / metabolism
  • HCT116 Cells
  • HeLa Cells
  • Humans
  • Mice
  • Myoblasts / metabolism
  • Nuclear Localization Signals / metabolism
  • Promoter Regions, Genetic
  • Protein Binding
  • Transfection

Substances

  • CCAAT-Binding Factor
  • Cell-Penetrating Peptides
  • DNA-Binding Proteins
  • NFYA protein, human
  • Nfya protein, mouse
  • Nuclear Localization Signals