title: Protein interaction screening identifies SH3RF1 as a new regulator of FAT1 protein levels.
abstract: Mutations and ectopic FAT1 cadherin expression are implicated in a broad spectrum of diseases ranging from developmental disorders to cancer. The regulation of FAT1 and its downstream signalling pathways remain incompletely understood. We hypothesized that identification of additional proteins interacting with the FAT1 cytoplasmic tail would further delineate its regulation and function. A yeast two-hybrid library screen carried out against the juxtamembrane region of the cytoplasmic tail of FAT1 identified the E3 ubiquitin-protein ligase SH3RF1 as the most frequently recovered protein-binding partner. Ablating SH3RF1 using siRNA increased cellular FAT1 protein levels and stabilized expression at the cell surface, while overexpression of SH3RF1 reduced FAT1 levels. We conclude that SH3RF1 acts as a negative post-translational regulator of FAT1 levels.
authors.affiliation: VIB Center for the Biology of Disease, Leuven, Belgium.|Hunter Cancer Research Alliance, University of Newcastle, Callaghan, Australia.
authors.forename: Charles E
authors.initials: CE
authors.lastname: de Bock
authors.affiliation: The Biomedical Research Centre, University of British Columbia, Vancouver, Canada.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Hughes
authors.affiliation: The Biomedical Research Centre, University of British Columbia, Vancouver, Canada.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Snyder
authors.affiliation: Hunter Cancer Research Alliance, University of Newcastle, Callaghan, Australia.|School of Environmental and Life Sciences, University of Newcastle, Callaghan, Australia.
authors.forename: Steven
authors.initials: S
authors.lastname: Alley
authors.affiliation: Hunter Cancer Research Alliance, University of Newcastle, Callaghan, Australia.|School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, Australia.
authors.forename: Elham
authors.initials: E
authors.lastname: Sadeqzadeh
authors.affiliation: Hunter Cancer Research Alliance, University of Newcastle, Callaghan, Australia.|School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, Australia.
authors.forename: Matt D
authors.initials: MD
authors.lastname: Dun
authors.affiliation: The Biomedical Research Centre, University of British Columbia, Vancouver, Canada.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: McNagny
authors.affiliation: The Kinghorn Cancer Centre and Cancer Research Program, Garvan Institute of Medical Research, Darlinghurst, Australia.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Molloy
authors.affiliation: Hunter Cancer Research Alliance, University of Newcastle, Callaghan, Australia.|School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, Australia.
authors.forename: Hubert
authors.initials: H
authors.lastname: Hondermarck
authors.affiliation: Hunter Cancer Research Alliance, University of Newcastle, Callaghan, Australia.|School of Environmental and Life Sciences, University of Newcastle, Callaghan, Australia.
authors.forename: Rick F
authors.initials: RF
authors.lastname: Thorne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D015820:Cadherins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D015870:Gene Expression; D006801:Humans; D047468:Immunoprecipitation; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D018909:src Homology Domains / Q000235:genetics
chemical_list: D015820:Cadherins; C502403:FAT1 protein, human; C498193:SH3RF1 protein, human; D044767:Ubiquitin-Protein Ligases
keywords: E3 ubiquitin-ligase; FAT1 cadherin; SH3RF1; protein half-life; protein-protein interaction; yeast two-hybrid