DOI

https://doi.org/10.25772/V74X-Y047

Defense Date

2019

Document Type

Thesis

Degree Name

Master of Science

Department

Molecular Biology and Genetics

First Advisor

Dr. Rita Shiang

Abstract

PNPase, polynucleotide phosphorylase, is a multifunctional exoribonuclease protein with 3` terminal oligonucleotide polymerase activity. Coded by the PNPT1 gene, the protein is associated with mitochondrial homeostasis and functions as a possible target for cancer therapy. In this study, C. elegans was used to investigate the effect of mutation and overexpression of pnpt-1, the gene that encodes PNPase. It was determined that two specific mutations in pnpt-1 did not affect PNPase expression nor did they produce deleterious phenotypes that affected polycistronic transcript accumulation or ROS production. Creation of a stable overexpression model was achieved through Fusion PCR. However, different transgenic strains overexpressing PNPase produced opposite results for polycistronic transcript accumulation while ROS production saw no significant change, suggesting a mosaic overexpression model. In a cancer model, exogenous PNPase was present in the pachytene region of the germline and where expressed the cells were in non-germline cells suggesting differentiation mechanisms associated with overexpression of PNPase. However, further analysis of different mutations in pnpt-1 or optimizations to the overexpression model are necessary to provide a better understanding of PNPase function with mitochondria homeostasis and in a cancer model setting.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

12-12-2019

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