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A novel peptide motif binding to and blocking the intracellular activity of the human papillomavirus E6 oncoprotein

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Abstract

Specific types of human papillomaviruses (HPVs) cause cervical cancer. The viral E6 oncogene is a critical factor for maintaining the malignant phenotype of HPV-positive tumour cells. By yeast two-hybrid screening of a randomised peptide expression library, we isolated linear short peptides, which specifically bind to the HPV16 E6 oncoprotein. Sequence alignments and mutational analyses of the peptides identified a hitherto undiscovered E6-binding motif. Intracellular expression of a peptide containing the novel E6-binding motif resulted in inhibition of colony formation capacity, specifically of HPV16-positive cancer cells. A solubility-optimised variant of the peptide was created, which binds to HPV16 E6 with high affinity. Its intracellular expression efficiently induced apoptosis in HPV16-positive cancer cells. This was linked to restoration of intracellular p53 activities. Thus, this newly identified E6-binding motif could form a novel basis for the development of rational strategies for the treatment of HPV16-positive preneoplastic and neoplastic lesions.

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Acknowledgements

This work was supported by the Deutsche Krebshilfe. We thank Angela Heilig for excellent technical assistance, Dr. H. Zentgraf for providing anti-hrGFP antibodies and Dr. Jennifer Reed for helpful discussions.

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Authors and Affiliations

  1. Molecular Therapy of Virus-Associated Cancers (F065), German Cancer Research Center, Im Neuenheimer Feld 242, 69120, Heidelberg, Germany

    Susanne Dymalla, Claudia Lohrey, Felix Hoppe-Seyler & Karin Hoppe-Seyler

  2. Department of Biology, University of Konstanz, Konstanz, Germany

    Martin Scheffner & Elvira Weber

  3. Chemical Biology Core Facility, EMBL, Heidelberg, Germany

    Peter Sehr

Authors
  1. Susanne Dymalla
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  2. Martin Scheffner
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  3. Elvira Weber
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  4. Peter Sehr
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  5. Claudia Lohrey
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  6. Felix Hoppe-Seyler
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  7. Karin Hoppe-Seyler
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Corresponding authors

Correspondence to Felix Hoppe-Seyler or Karin Hoppe-Seyler.

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Fig. S1

Total blots obtained by Western blot and used for the indicated figures. Dashed lines indicate where membranes were cut to allow for immunoprinting of a single membrane with different antibodies, as detailed aside. For experimental design, see legends of the indicated figure and methods. a Full scan for Fig. 7a. b Full scan for Fig. 7b. Bold lines indicate which lanes were cut and joined for Fig. 7b. Tubulin immunoblot marked with * was included in Fig. 7b. (PDF 5.03 KB)

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Dymalla, S., Scheffner, M., Weber, E. et al. A novel peptide motif binding to and blocking the intracellular activity of the human papillomavirus E6 oncoprotein. J Mol Med 87, 321–331 (2009). https://doi.org/10.1007/s00109-008-0432-1

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  • DOI: https://doi.org/10.1007/s00109-008-0432-1

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