Date of Award
2016
Embargo Period
8-1-2024
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.
Recommended Citation
Karam, Joseph A., "Hop, The Heat Shock Link between Hsp90 and the Piwi-piRNA Pathway" (2016). MUSC Theses and Dissertations. 401.
https://medica-musc.researchcommons.org/theses/401
Rights
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