title: Characterization of nuclear PTEN and its post translational modifications.
abstract: Somatic loss-of-function mutations of PTEN are found in a variety of human malignancies. Our recent work demonstrated that the nuclear function of PTEN is implicated in the maintenance of genome integrity. Proper subcellular localization of PTEN following genotoxic stress is coordinated by a cellular mechanism that involves post-translational modification by SUMOylation and ATM-mediated phosphorylation. Here we summarize biochemical and cell-based methodologies that can be used to characterize the SUMOylation and phosphorylation state of nuclear PTEN in the context of DNA damage. In addition, we describe assays to determine the biological function of SUMO-PTEN in homologous recombination DNA repair. These methods will help elucidate the precise molecular mechanisms of PTEN's role in the maintenance of genomic stability.
authors.affiliation: Princess Margaret Cancer Center, University Health Network, Toronto, Ontario M5G 2M9, Canada.
authors.forename: J
authors.initials: J
authors.lastname: Ho
authors.affiliation: Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Bassi
authors.affiliation: Princess Margaret Cancer Center, University Health Network, Toronto, Ontario M5G 2M9, Canada; Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada. Electronic address: vuks@uhnres.utoronto.ca.
authors.forename: V
authors.initials: V
authors.lastname: Stambolic
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009154:Mutation / Q000502:physiology; D051059:PTEN Phosphohydrolase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D058207:Sumoylation / Q000502:physiology*; D025521:Tumor Suppressor Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
chemical_list: D025521:Tumor Suppressor Proteins; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human
keywords: ATM; DNA damage; PTEN; Phosphorylation; SUMOylation