title: A biosensor to monitor dynamic regulation and function of tumour suppressor PTEN in living cells.
abstract: Tumour suppressor PTEN is a phosphatase that negatively regulates the PI3K/AKT pathway. The ability to directly monitor PTEN conformation and function in a rapid, sensitive manner is a key step towards developing anti-cancer drugs aimed at enhancing or restoring PTEN-dependent pathways. Here we developed an intramolecular bioluminescence resonance energy transfer (BRET)-based biosensor, capable of detecting signal-dependent PTEN conformational changes in live cells. The biosensor retains intrinsic properties of PTEN, enabling structure-function and kinetic analyses. BRET shifts, indicating conformational change, were detected following mutations that disrupt intramolecular PTEN interactions, promoting plasma membrane targeting and also following physiological PTEN activation. Using the biosensor as a reporter, we uncovered PTEN activation by several G protein-coupled receptors, previously unknown as PTEN regulators. Trastuzumab, used to treat ERBB2-overexpressing breast cancers also elicited activation-associated PTEN conformational rearrangement. We propose the biosensor can be used to identify pathways regulating PTEN or molecules that enhance its anti-tumour activity.
authors.affiliation: 1] Department of Endocrinology, Metabolism and Diabetes, Inserm, U1016, Institut Cochin, 27 rue du Faubourg St Jaques, Paris 75014, France [2] CNRS, UMR8104, Paris 75014, France [3] University Paris Descartes, Sorbonne Paris Cité, Paris 75014, France [4].
authors.forename: Evelyne
authors.initials: E
authors.lastname: Lima-Fernandes
authors.affiliation: 1] Department of Endocrinology, Metabolism and Diabetes, Inserm, U1016, Institut Cochin, 27 rue du Faubourg St Jaques, Paris 75014, France [2] CNRS, UMR8104, Paris 75014, France [3] University Paris Descartes, Sorbonne Paris Cité, Paris 75014, France [4].
authors.forename: Stanislas
authors.initials: S
authors.lastname: Misticone
authors.affiliation: 1] Department of Endocrinology, Metabolism and Diabetes, Inserm, U1016, Institut Cochin, 27 rue du Faubourg St Jaques, Paris 75014, France [2] CNRS, UMR8104, Paris 75014, France [3] University Paris Descartes, Sorbonne Paris Cité, Paris 75014, France [4].
authors.forename: Cédric
authors.initials: C
authors.lastname: Boularan
authors.affiliation: 1] Molecular Biology Program, Department of Biochemistry, Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, Quebec, Canada H3C 3J7 [2] Department of Pathology and Cell Biology, Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, Quebec, Canada H3C 3J7 [3].
authors.forename: Justine S
authors.initials: JS
authors.lastname: Paradis
authors.affiliation: 1] Department of Endocrinology, Metabolism and Diabetes, Inserm, U1016, Institut Cochin, 27 rue du Faubourg St Jaques, Paris 75014, France [2] CNRS, UMR8104, Paris 75014, France [3] University Paris Descartes, Sorbonne Paris Cité, Paris 75014, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Enslen
authors.affiliation: 1] Molecular Biology Program, Department of Biochemistry, Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, Quebec, Canada H3C 3J7 [2] Department of Pathology and Cell Biology, Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, Quebec, Canada H3C 3J7.
authors.forename: Philippe P
authors.initials: PP
authors.lastname: Roux
authors.affiliation: 1] Molecular Biology Program, Department of Biochemistry, Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, Quebec, Canada H3C 3J7 [2] Department of Pathology and Cell Biology, Institute for Research in Immunology and Cancer, Université de Montréal, Montréal, Quebec, Canada H3C 3J7.
authors.forename: Michel
authors.initials: M
authors.lastname: Bouvier
authors.affiliation: Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
authors.forename: George S
authors.initials: GS
authors.lastname: Baillie
authors.affiliation: 1] Department of Endocrinology, Metabolism and Diabetes, Inserm, U1016, Institut Cochin, 27 rue du Faubourg St Jaques, Paris 75014, France [2] CNRS, UMR8104, Paris 75014, France [3] University Paris Descartes, Sorbonne Paris Cité, Paris 75014, France.
authors.forename: Stefano
authors.initials: S
authors.lastname: Marullo
authors.affiliation: 1] Department of Endocrinology, Metabolism and Diabetes, Inserm, U1016, Institut Cochin, 27 rue du Faubourg St Jaques, Paris 75014, France [2] CNRS, UMR8104, Paris 75014, France [3] University Paris Descartes, Sorbonne Paris Cité, Paris 75014, France.
authors.forename: Mark G H
authors.initials: MG
authors.lastname: Scott
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: D051059:PTEN Phosphohydrolase; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human; C494929:PTEN protein, human