Date of Award

2014

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Committee Chair

Roy Magnuson

Committee Member

Gordon MacGregor

Committee Member

Luis R. Cruz-Vera

Subject(s)

Bacteriophages--Genetics, Bacterial genetics, Phosphorylation, Toxins, Antitoxins, Protein kinases

Abstract

Doc induces cell death by inhibiting translation; however, the mechanism of Doc-induced cell death and the cellular target of the toxin were unknown. One theory suggested that Doc inhibits translation elongation by binding directly to the 30S ribosomal subunit. Later evidence showed catalytic activity in distant homologs of Doc. These homologs contain a Fic-domain that has been shown to modify target GTPases by AMPylation and phosphocholination. Therefore, [35S] - Met, á[32P] - ATP, and ã[32P] - ATP were used in conjunction with an S30 extract to confirm that Doc inhibits translation, to assess the mechanism of modification, and to identify the modified target. The results showed that Doc is an enzyme that inhibits translation and phosphorylates a protein target.

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