Publication Date

2014

Document Type

Thesis

Committee Members

Katherine Excoffon (Advisor), Lynn Hartzler (Committee Member), Mill Miller (Committee Member)

Degree Name

Master of Science (MS)

Abstract

Adenovirus's primary receptor, the Coxsackievirus and adenovirus receptor (CAR), has two transmembrane isoforms with differential localization within polarized epithelial cells (CAREx7 (basolateral), CAREx8 (apical)). I hypothesized that each isoform of CAR is degraded at different rates and is regulated by Src-family kinases. Increasing or decreasing the concentration of CAR is predicted to directly alter adenoviral entry. To test this hypothesis, inducible Madin Darby Canine Kidney (MDCK) cell lines either expressing CAREx7, CAREx8, or mCherry, under a doxycycline-inducible promoter were used in pulse-chase structured experiments to calculate the half-lives of these proteins. Alternatively, a polarized model lung epithelium (CaLu-3) treated with Src-kinase inhibitor, PP2, was used to test apical CAREx8 expression and viral transduction. Though CAREx7 and CAREx8 did have varying half-lives (5.1 h, 2.0 h respectively), inhibiting Src-family kinases did not decrease CAREx8 expression at early time points but did diminish adenovirus transduction. These data confirm that Src does not play a role in CAR turnover, but remains critical for adenoviral infection.

Page Count

112

Department or Program

Department of Biological Sciences

Year Degree Awarded

2014

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.


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