Undergraduate Honors Theses

Thesis Defended

Spring 2019

Document Type


Type of Thesis

Departmental Honors


Molecular, Cellular, & Developmental Biology

First Advisor

Robert Garcea, MD


Murine Polyomaviruses are dsDNA viruses that hijack the host’s DNA damage response (DDR) pathway to replicate their own genomes (Heiser et al., 2016), offering a model for human polyomavirus replication. The viral protein large T-antigen (LT) is essential for viral replication and can interact with a variety of DDR proteins in viral DNA replication centers (Brodsky & Pipas, 1998). Replication protein A (RPA) is a DDR protein complex made up of three subunits, RPA70, RPA32, and RPA14. RPA70 and RPA32 have been shown to directly interact with LT in cells over-expressing each protein, either individually or together (Banerjee et al., 2013). Individual point mutations within LT could disrupt the LT-RPA interaction (Banerjee et al., 2013). However, the LT–RPA interaction has yet to be explored in the context of viral infection. This study aimed to evaluate mCherry-tagged RPA32 binding with wild-type LT and previously characterized point-mutants E320A and K308E during infection of mouse fibroblasts (Banerjee et al., 2013). We show that, vDNA was replicated from genomes encoding both wild-type and mutant LT proteins. However, we did not observe an interaction between wild-type LT and mCherry-tagged mouse RPA32 (mCh-muRPA32).