G-quadruplex DNA recognition by nucleophosmin: new insights from protein dissection

Biochim Biophys Acta. 2014 Jun;1840(6):2050-9. doi: 10.1016/j.bbagen.2014.02.017. Epub 2014 Feb 24.

Abstract

Background: Nucleophosmin (NPM1, B23) is a multifunctional protein that is involved in a variety of fundamental biological processes. NPM1/B23 deregulation is implicated in the pathogenesis of several human malignancies. This protein exerts its functions through the interaction with a multiplicity of biological partners. Very recently it is has been shown that NPM1/B23 specifically recognizes DNA G-quadruplexes through its C-terminal region.

Methods: Through a rational dissection approach of protein here we show that the intrinsically unfolded regions of NPM1/B23 significantly contribute to the binding of c-MYC G-quadruplex motif. Interestingly, the analysis of the ability of distinct NPM1/B23 fragments to bind this quadruplex led to the identifications of distinct NPM1/B23-based peptides that individually present a high affinity for this motif.

Results: These results suggest that the tight binding of NPM1/B23 to the G-quadruplex is achieved through the cooperation of both folded and unfolded regions that are individually able to bind it. The dissection of NPM1/B23 also unveils that its H1 helix is intrinsically endowed with an unusual thermal stability.

Conclusions: These findings have implications for the unfolding mechanism of NPM1/B23, for the G-quadruplex affinity of the different NPM1/B23 isoforms and for the design of peptide-based molecules able to interact with this DNA motif.

General observation: This study sheds new light in the molecular mechanism of the complex NPM1/G-quadruplex involved in acute myeloid leukemia (AML) disease.

Keywords: Disordered protein region; Helical stability; Surface Plasmon Resonance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • G-Quadruplexes*
  • Humans
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / physiology*
  • Nucleophosmin
  • Protein Folding
  • Proto-Oncogene Proteins c-myc / metabolism

Substances

  • MYC protein, human
  • NPM1 protein, human
  • Nuclear Proteins
  • Proto-Oncogene Proteins c-myc
  • Nucleophosmin