Date of Award
Summer 8-6-2020
Degree Type
Thesis
Degree Name
MS Microbiology
Department
Biology
Advisor
Daniel Brian Nichols, Ph.D.
Committee Member
Tinchun Chu, Ph.D.
Committee Member
Jane Ko, Ph.D.
Keywords
Molluscum Contagiosum Virus, MCV, Yeast Two-Hybrid, MC159, BAP29
Abstract
The human immune system acts as our primary defense against invading pathogens including bacteria, parasites, and viruses. Molluscum Contagiosum Virus (MCV) is a poxvirus that causes persistent skin lesions in humans that can last for many months at a time. Persistence of MCV infections is attributed in part to the ability of the virus to suppress human immune signaling. Many MCV genes, including MC159, MC160, and MC163, code for products that inhibit several important innate immune signaling pathways and consequently reduce inflammation, leukocyte infiltration, and chemokine production. However, due to lack of a tissue culture system in which to propagate MCV, many suspected MCV immune evasion genes remain understudied or not studied in the slightest. Further, the molecular mechanisms of known MCV immune evasion products require further elucidation. Thus, this study has set forth to establish a yeast two-hybrid system to discover unknown protein-protein interactions between MCV proteins and a library of human proteins. MCV genes were inserted inside of bait vectors for use in the system. Given it is the most extensively studied gene to date, MC159 was chosen first to be transformed into yeast and ran through the system against a library of human protein prey vectors. This experiment showed success in the form of dozens of positive hits, six of which were later sequenced and identified through bioinformatic techniques. One of these hits, BAP29, was of special interest due to its suspected influence on immune signaling, specifically trafficking of MHC I out of the endoplasmic reticulum. This is the first reported interaction between MC159 and BAP29 and may represent a novel interaction. While further experiments are needed to confirm this, these results show that the yeast two-hybrid assay may be an effective method for discovering individual MCV genes’ human protein targets.
Recommended Citation
Bryan, Eric, "Developing a Yeast Two-Hybrid System to Study Molluscum Contagiosum Virus Protein-Protein Interactions with Human Proteins" (2020). Seton Hall University Dissertations and Theses (ETDs). 2805.
https://scholarship.shu.edu/dissertations/2805
