Date of Award

12-31-2015

Document Type

Campus Access Thesis

Degree Name

Master of Science (MS)

Department

Biotechnology and Biomedical Science

First Advisor

Linda Huang

Second Advisor

Alexey Veraksa

Third Advisor

Michael Shiaris

Abstract

In S. cerevisiae, the MAP kinase encoded by SMK1 regulates sporulation. My work examines Smk1 localization and identifies and characterizes new protein interactors of Smk1. We see Smk1 localizing along the growing prospore membrane (PSM) and at the leading edge of the PSM. Using mass spectrometry analysis, two sets of potential protein interactions were identified that occur when Smk1 is localized to either the PSM or the leading edge. This work focuses on two proteins identified within these data sets: Ssp2 and Isc10. We see that Ssp2 localizes along the PSM and to the leading edge, similar to Smk1, and that relocation of Smk1 to the leading edge is dependent on SSP2. Additionally, Ssp2 and Smk1 are shown to physically interact, and that the activation state of Smk1 has no effect on its interaction with Ssp2. By examining the role of ISC10 during sporulation, we see that Isc10 localizes along the PSM as distinct puncta and that Isc10 expression is induced 6 hours into sporulation. Isc10 undergoes a post-translation modification, which is dependent on SMK1. Late during spore development, spores lacking ISC10 develop abnormally large vacuoles, suggesting that ISC10 may regulate vacuolar proliferation.

Comments

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