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Date Submitted

Spring 5-1-2013

Research Mentor and Department

Dr. Douglas Chalker

Restricted/Unrestricted

Dissertation/Thesis

Abstract

Tetrahymena is part of a group of eukaryotes known as ciliates. These organisms have many unique qualities such as rows of cilia and nuclear dimorphism that make them an interesting group with which to work. Tetrahymena has specifically been used as model organism because of its short generation times, ease of transduction, as well as its relatively large cell-size, which facilitates visualization. The ciliated structure as well as the functionally distinct nuclei facilitated study of the localization and function of both of these proteins because STK1 serves as a cilia production regulator, while Sub1 acts a transcriptional co-activator of RNA polymerase II. Construction of YFP-fusion proteins allowed visualization of the localization by fluorescence microscopy. Through these methods, STK1 was seen to localize around the basal bodies found in the epiplasm. These results were reasonable considering STK1’s function in ciliary production, which occurs at basal bodies. Sub1 localized to the macronucleus; this was logical since the macronucleus is the transcriptionally active nucleus. Both proteins are upregulated during conjugation, which was shown through RT-PCR. Research in these proteins can facilitate translational studies. The cilia structure of Tetrahymena is used to model lung cilia, and so further research about STK1 could prove beneficial in understanding the mechanisms surrounding human lung cilia function. Further research about Sub1 could enhance understanding of gene regulation and activation because Sub1 contains a conserved PC4 domain, which is known to activate gene transcription in mammals.