Date of Award:

5-1-2003

Document Type:

Thesis

Degree Name:

Master of Science (MS)

Department:

Biology

Committee Chair(s)

Daryll B. DeWald

Committee

Daryll B. DeWald

Committee

Jon Y. Takemoto

Committee

Marie K. Walsh

Abstract

Phosphoinositides are membrane-bound lipids that exist as eight different isoforms resulting from the phosphorylation of their inositol head group at three distinct positions. Proteins with Pleckstrin Homology (PH) domains bind to phosphoinositides involved in the regulation of cellular processes. The binding of several yeast PH domains to phosphoinositides was examined using dot-blot overlay assays, Surface Plasmon Resonance, and in vivo localization methodologies. While most of these yeast PH domains exhibited low specificity and/or low binding affinity, several proteins showed strong in vivo localization to either the plasma membrane or the Golgi membrane in mammalian cells. The role of the YPR115 gene was also studied by examining the growth defects of a knockout strain and a potential dominant negative strain compared to the parental strain on SD-ura plates exposed to 100 mM, 250 mM, and 500 mM NaCl. In addition, growth of the YPR115 mutant strains at 30°C and 37°C was documented on SD-ura plates and in liquid media. The knockout strain exhibited a slightly smaller colony size with NaCl plates and minimal growth was detected at 37°C. The potential dominant negative strain showed no discernable growth defect compared to the parental strain with NaCl at 37°C.

Included in

Biology Commons

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