β-catenin is O-GlcNAc modified at Serine 23: Implications for β-catenin’s Subcellular Distribution and Transcriptional Activity

  • Author / Creator
    Ha, Jacqueline R
  • β-catenin is a potent oncoprotein that serves as a structural anchor at the adherens junctions and as a transcriptional co-activator of the Wnt Signaling pathway. β-catenin was identified to be post-translationally modified by O-linked β-D-N-acetyl-glucosamine (O-GlcNAc). This investigation was aimed to identify Serine 23 (Ser23) as a site for O-GlcNAc modification and to characterize the relevance of this site for β-catenin’s function.
    Serine 23 to Glycine mutant or wild-type β-catenin constructs were expressed in DU145 cell line and subsequently treated with PUGNAc—a drug that globally increases O-GlcNAcylation. O-GlcNAc-β-catenin levels were characterized by Wheat Germ Agglutinin (WGA)-HRP or WGA-agarose precipitation. Alterations in β-catenin’s subcellular localization, interactions, and transcriptional activity were analyzed through confocal microscopy, immunoprecipitation, and, RT-qPCR and luciferase reporter assay, respectively.
    This study demonstrated that Ser23 of β-catenin was a site for O-GlcNAcylation which increased β-catenin’s localization to the plasma membrane and decreased its transcriptional activity within the nucleus.

  • Subjects / Keywords
  • Graduation date
    Fall 2012
  • Type of Item
  • Degree
    Master of Science
  • DOI
  • License
    This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.
  • Language
  • Institution
    University of Alberta
  • Degree level
  • Department
  • Specialization
    • Medical Sciences-Paediatrics
  • Supervisor / co-supervisor and their department(s)
  • Examining committee members and their departments
    • Pasdar, Manijeh (Cell Biology)
    • Baksh, Shairaz (Paediatrics)
    • Moore, Ronald (Surgery)