Date of Award

2016

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biochemistry and Molecular Biology

College

College of Graduate Studies

First Advisor

Vamsi Gangaraju

Second Advisor

Tilman Heise

Third Advisor

Julie Woolworth-Hirschhorn

Fourth Advisor

Je Hyun Yoon

Fifth Advisor

David Long

Abstract

The Piwi-piRNA pathway is an RNA interference pathway that is conserved across eukaryotes; it is involved with the maintenance of genomic stability through its actions on the germline. We have studied the relationship between a protein complex involving factors in the Piwi-piRNA pathway, Piwi and Aubergine, and the Hsp90-Hop-Hsp70 molecular chaperone machinery. Through a series of in vitro experiments and previous data, we have shown that an interaction between Piwi, Hop, and Hsp90 occurs, when all three proteins are present. Shedding light on this interaction could give us insight on how the PiwipiRNA pathway functions. Through in vivo experiments performed, we have shown that germline knockdowns of Hop, Hsc70-4, and Hsp83 have impacts on the Piwi-piRNA pathway’s ability to perform its function suppressing transposable elements. Through deep sequencing of the total small RNA population present under a Hop germline knockdown, as well as analysis of the population of piRNAs bound to Piwi when Hop is knocked down, we have shown the first evidence of molecular chaperone involvement in both the biogenesis and the loading of piRNAs onto Piwi. This work will help to elucidate the function of the Piwi-piRNA pathway in Drosophila melanogaster. We aim to continue this work by performing deep sequencing experiments on the Hsc70-4 and Hsp83 germline knockdown samples to fully assess their impacts on the loading and biogenesis of piRNAs.

Rights

All rights reserved. Copyright is held by the author.

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