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Theses and Dissertations

Characterization of Arf4•GDP Open Access

Descriptions

Other title
Subject/Keyword
dominant
puncta
peripheral
Arf
negative
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Summerfeldt, Nathan
Supervisor and department
Melançon, Paul (Cell Biology)
Examining committee member and department
Glover, Mark (Biochemistry)
LaPointe, Paul (Cell Biology)
Simmen, Thommas (Cell Biology)
Department
Department of Cell Biology
Specialization

Date accepted
2010-07-15T16:08:54Z
Graduation date
2010-11
Degree
Master of Science
Degree level
Master's
Abstract
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.
Language
English
Rights
License granted by Nathan Summerfeldt (summerfe@ualberta.ca) on 2010-07-14T16:30:23Z (GMT): Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of the above terms. The author reserves all other publication and other rights in association with the copyright in the thesis, and except as herein provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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File author: Nathan
Page count: 111
File language: en-CA
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