DARPin-induced reactivation of p53 in HPV-positive cells

Philipp Münick; Alexander Strubel; Dimitrios-Ilias Balourdas; Julianne S Funk; Marco Mernberger; Christian Osterburg; Birgit Dreier; Jonas V Schaefer; Marcel Tuppi; Büşra Yüksel; Birgit Schäfer; Stefan Knapp; Andreas Plückthun; Thorsten Stiewe; Andreas C Joerger; Volker Dötsch

doi:10.1038/s41594-024-01456-7

DARPin-induced reactivation of p53 in HPV-positive cells

Nat Struct Mol Biol. 2025 Jan 9. doi: 10.1038/s41594-024-01456-7. Online ahead of print.

Authors

Philipp Münick¹, Alexander Strubel¹, Dimitrios-Ilias Balourdas^{2

3}, Julianne S Funk⁴, Marco Mernberger⁴, Christian Osterburg¹, Birgit Dreier⁵, Jonas V Schaefer⁵, Marcel Tuppi¹, Büşra Yüksel^{1

6}, Birgit Schäfer¹, Stefan Knapp^{2

3}, Andreas Plückthun⁵, Thorsten Stiewe^{4

7

8}, Andreas C Joerger^{2

3}, Volker Dötsch⁹

Affiliations

¹ Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, Goethe University, Frankfurt, Germany.
² Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany.
³ Structural Genomics Consortium, Goethe University, Frankfurt, Germany.
⁴ Institute of Molecular Oncology, Universities of Giessen and Marburg Lung Center, Member of the German Center for Lung Research, Philipps-University, Marburg, Germany.
⁵ Department of Biochemistry, University of Zurich, Zurich, Switzerland.
⁶ IMPRS on Cellular Biophysics, Frankfurt, Germany.
⁷ Genomics Core Facility, Philipps-University, Marburg, Germany.
⁸ Institute for Lung Health, Justus Liebig University, Giessen, Germany.
⁹ Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance, Goethe University, Frankfurt, Germany. [email protected].

PMID: 39789211
DOI: 10.1038/s41594-024-01456-7

Abstract

Infection of cells with high-risk strains of the human papillomavirus (HPV) causes cancer in various types of epithelial tissue. HPV infections are responsible for ~4.5% of all cancers worldwide. Tumorigenesis is based on the inactivation of key cellular control mechanisms by the viral proteins E6 and E7. The HPV E6 protein interacts with the cellular E3 ligase E6AP, and this complex binds to the p53 DNA-binding domain, which results in degradation of p53. Inhibition of this interaction has the potential to reactivate p53, thus preventing oncogenic transformation. Here we describe the characterization of a designed ankyrin repeat protein that binds to the same site as the HPV E6 protein, thereby displacing the E3 ligase and stabilizing p53. Interaction with the designed ankyrin repeat protein does not affect p53 DNA binding or the crucial MDM2 negative feedback loop but reactivates a p53-dependent transcriptional program in HeLa (HPV18-positive) and SiHa (HPV16-positive) cells, suggesting a potential therapeutic use.