Author

Darren Ferree

Date of Award

2015

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Committee Chair

Roy D. Magnuson

Committee Member

Luis R. Cruz-Vera

Committee Member

Gordon MacGregor

Subject(s)

Plasmids., Molecular genetics., Bacteriophages--Genetics., Phosphorylation., Protein kinases.

Abstract

Doc toxin from the bacteriophage P1 plasmid addiction operon kills daughter cells that are cured of plasmid. At the beginning of this project it was known that Doc is a kinase that transfers the phosphoryl group from the gamma position of ATP to some unidentified target protein of approximately 45 kDa molecular weight. To better characterize this target, assays were conducted to determine optimal conditions for Doc-dependent phosphorylation of target, and these conditions were then used to generate samples for analysis by two-dimensional polyacrylamide gel electrophoresis, liquid chromatography-coupled tandem mass spectrometry, and immunoprecipitation. The results of these experiments indicate that some particular species of bacterial Elongation Factor Tu (EF-Tu) is the target of Doc but that not all species of EF-Tu are readily phosphorylated by Doc.

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