Characterization of Arf4•GDP

  • Author / Creator
    Summerfeldt, Nathan
  • In this thesis, I characterized the association of Arf4•GDP with ER-Golgi intermediate compartment membranes. We confirmed that GDP-arrested Arf4 mutants associated with membranes irrespective of nature of tag or mutation. Recruitment appeared specific since loss of N-terminal myristoylation abolished binding. Surprisingly, mutations of residues unique to class II Arfs did not prevent recruitment of Arf4 to peripheral puncta. We then examined the failure of the GDP-arrested Arf4 mutant to disrupt Golgi structure. We identified residues R79 and E113 (likely involved in salt bridge interaction) only present in Arf1 and Arf5 as critical to the ability of their GDP-arrested mutants to disrupt Golgi structure. As predicted, introduction of these residues transformed Arf4•GDP into a dominant negative mutant. Interestingly, overexpression of the putative Arf•GDP receptor membrin prevented the effects of dominant negative Arf1 but not dominant negative Arf4. These results will facilitate identification of a novel Arf target critical to protein trafficking.

  • Subjects / Keywords
  • Graduation date
    Fall 2010
  • 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
  • Supervisor / co-supervisor and their department(s)
  • Examining committee members and their departments
    • LaPointe, Paul (Cell Biology)
    • Simmen, Thommas (Cell Biology)
    • Glover, Mark (Biochemistry)