Date of Award

Fall 2019

Document Type

Thesis Restricted

Degree Name

Master of Science (MS)

Department

Biology

Committee Chairperson

Erin Gestl, Ph.D.

Committee Member

Jessica Sullivan-Brown, Ph.D.

Committee Member

Gustave Mbuy, Ph.D.

Abstract

Cells replicate their genome efficiently and effectively via a combination of numerous DNA polymerases. Several polymerases perform specific functions in the realm of DNA damage repair and translesion synthesis, these functions are aimed at increasing cellular survivability in the face of genomic damage. Polymerase Zeta, a translesion synthesis polymerase, is has been studied in a multitude of model systems with differing observations. In yeast, knockdown of Pol Zeta leads to increased genomic stability and results in total embryonic lethality in mice but there have been no observations made yet in zebrafish. This study seeks to examine this polymerase in the Zebrafish model system by knocking down its catalytic subunit, Rev3, with morpholino oligomers. To assess the impact of the knockdown, the comet assay and an acridine orange assay will be conducted to measure apoptosis caused by DNA damage induced by UV treatment. Our data suggests that the removal of Pol Zeta results in an increase of apoptosis higher that of a control group that did not undergo injection of the morpholinos. This finding highlights Pol Zeta as an essential survivability polymerase when the genome is faced with multiple lesion sites during replication. These results lay a groundwork for further translesion synthesis polymerase knockdown experiments separate from, or in addition to knocking down Pol Zeta. The accessory subunit of Pol Zeta, Rev7, is a target of future research due to its roles with the creation of the Pol Zeta complex.

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