title: Targeting the Two Oncogenic Functional Sites of the HPV E6 Oncoprotein with a High-Affinity Bivalent Ligand.
abstract: The E6 oncoproteins of high-risk mucosal (hrm) human papillomaviruses (HPVs) contain a pocket that captures LxxLL motifs and a C-terminal motif that recruits PDZ domains, with both functions being crucial for HPV-induced oncogenesis. A chimeric protein was built by fusing a PDZ domain and an LxxLL motif, both known to bind E6. NMR spectroscopy, calorimetry and a mammalian protein complementation assay converged to show that the resulting PDZ-LxxLL chimera is a bivalent nanomolar ligand of E6, while its separated PDZ and LxxLL components are only micromolar binders. The chimera binds to all of the hrm-HPV E6 proteins tested but not to low-risk mucosal or cutaneous HPV E6. Adenovirus-mediated expression of the chimera specifically induces the death of HPV-positive cells, concomitant with increased levels of the tumour suppressor P53, its transcriptional target p21, and the apoptosis marker cleaved caspase 3. The bifunctional PDZ-LxxLL chimera opens new perspectives for the diagnosis and treatment of HPV-induced cancers.
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Juan
authors.initials: J
authors.lastname: Ramirez
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Juline
authors.initials: J
authors.lastname: Poirson
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Clémence
authors.initials: C
authors.lastname: Foltz
authors.affiliation: IGBMC, 1 rue Laurent Fries, BP 10142, 67404 Illkirch (France).
authors.forename: Yassmine
authors.initials: Y
authors.lastname: Chebaro
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Maxime
authors.initials: M
authors.lastname: Schrapp
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Amandine
authors.initials: A
authors.lastname: Meyer
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Anaëlle
authors.initials: A
authors.lastname: Bonetta
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Anne
authors.initials: A
authors.lastname: Forster
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Virus à ARN, Département de Virologie & UMR CNRS 3569, Paris (France).
authors.forename: Yves
authors.initials: Y
authors.lastname: Jacob
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: Murielle
authors.initials: M
authors.lastname: Masson
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France).
authors.forename: François
authors.initials: F
authors.lastname: Deryckère
authors.affiliation: Oncoprotein Team, Équipe Labellisée Ligue 2015, UMR CNRS-UDS 7242, École Supérieure de Biotechnologie de Strasbourg, Boulevard Sébastien Brant, BP 10413, F-67412 Illkirch (France). trave@unistra.fr.
authors.forename: Gilles
authors.initials: G
authors.lastname: Travé
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D020816:Amino Acid Motifs; D001665:Binding Sites; D016923:Cell Death; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression; D006367:HeLa Cells; D052162:Human papillomavirus 16 / Q000737:chemistry / Q000378:metabolism*; D052161:Human papillomavirus 18 / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D009369:Neoplasms / Q000378:metabolism / Q000628:therapy / Q000821:virology*; D009856:Oncogene Proteins, Viral / Q000737:chemistry / Q000378:metabolism*; D054731:PDZ Domains; D030361:Papillomavirus Infections / Q000378:metabolism / Q000628:therapy / Q000821:virology*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C068651:E6 protein, Human papillomavirus type 16; C052603:E6 protein, Human papillomavirus type 18; D008024:Ligands; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D016159:Tumor Suppressor Protein p53
keywords: antitumor agents; human papillomavirus; inhibitors; peptides; protein engineering