Publication Date

2023

Document Type

Thesis

Committee Members

Abimbola O. Kolawole, Ph.D. (Committee Co-Chair); Dawn P. Wooley, Ph.D. (Committee Co-Chair); Nancy J. Bigley, Ph.D. (Committee Member)

Degree Name

Master of Science (MS)

Abstract

Adenovirus is a human pathogen that causes cold like symptoms in healthy individuals, which could be lethal in pediatrics and the immunocompromised. In polarized epithelia, adenovirus uses the eight-exon isoform of the Coxsackievirus and Adenovirus Receptor (CAREx8) on the apical surface, to facilitate infection. CAREx8 protein expression level is regulated by the scaffolding protein MAGI-1, using two of its PSD95/Dlg-1/ZO-1 (PDZ) domains: PDZ2 and PDZ4. We previously showed that CAREx8 is degraded after interaction with PDZ4 and rescued following interaction with PDZ2. We then developed a PDZ2 binding decoy molecule (E6) that blocks the interaction between CAREx8 and PDZ2, which increases degradation of CAREx8 and, consequently, decreases adenovirus infection. We then synthesized TAT, a cell penetrating peptide (CPP) to the N-terminus of E6 for easy delivery into cells. In comparison to a scrambled control peptide, TAT_E6 significantly reduced CAREx8 expression and adenovirus-5 transduction in MDCK cells that stably expressed human CAREx8. However, recent studies suggest that there may be other CPPs that when conjugated to E6, will decrease CAREx8 protein expression and subsequently AdV transduction with higher efficacy than TAT_E6. As such, I selected five new CPPs, conjugated them to E6, then tested their CAREx8 protein expression reducing capabilities via Western blot analysis. These results demonstrated that all five of the selected E6-conjugated CPPs reduced CAREx8 protein expression more significantly than TAT_E6.

Page Count

62

Department or Program

Microbiology and Immunology

Year Degree Awarded

2023


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