Novel use of a cell-penetrating peptide-adaptor system to characterize Akirin constructs and investigate activity of bacterial effector proteins
Date of Award
Master of Science in Integrative Biology (MSIB)
First Committee Member
Second Committee Member
Third Committee Member
In the late 1980s, HIV protein TAT was found to confer the passage of HIV through the plasma membrane of mammalian cells. The McMurry lab developed a CPP-adaptor system which consists of the fusion of TAT to calmodulin to which mediates the delivery of exogenous proteins into cells and prevents endosomal entrapment. Calmodulin (CaM) is capable of binding to the calmodulin binding site (CBS) of cargo only in the presence of calcium. TAT-CaM can be used to characterize Akirin constructs and investigate the activity of bacterial effectors, AopP and AvrA, in the NF-kB pathway. Akirin is a small nuclear protein that was shown to interact with transcription factor Twist and chromatin remodeler Brahma. The functional regions of Akirin are unknown; therefore, TAT-CaM was used to characterize the four conserved regions of Akirin. Bacterial effectors, AopP and AvrA, have been shown to inhibit the NF-kB pathway. Previous studies used infection and transfection methods to study their activity. TAT-CaM provides a method that is non-toxic and physiologically applicable to study their activity. Based on this study, the binding kinetics between TAT-CaM and these proteins are successful, as well as TAT-CaM’s ability to mediate their delivery into mammalian cells. However, a more in-depth characterization of the Akirin regions and the activity of the bacterial effectors are still under investigation.
Available for download on Monday, June 26, 2023