Event Title
Effects of Cardiac Arrhythmic Mutant D95V on Calmodulin Structure and Stability
Location
CSU Ballroom
Start Date
11-4-2017 10:00 AM
End Date
11-4-2017 11:30 AM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Allison Land
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
APOBEC3A is a catalytically active DNA cytosine deaminase expressed in monocyte immune cells. This function allows APOBEC3A to mutate and restrict viruses, potentially including HIV. HIV-1, the causative agent of the major HIV/AIDS pandemic, is incapable of infecting monocytes. HIV-2, a less common variant, is capable of infecting monocytes. The unique protein Vpx, produced by HIV-2, but not HIV-1, is thought to be responsible for allowing HIV-2 infection in this immune cell. The objective of this study is to determine the sensitivity of human APOBEC3A to HIV-2 Vpx. We hypothesize that HIV-2 Vpx will be capable of mediating degradation of APOBEC3A and limiting its mutagenic capabilities, thus allowing HIV-2 to infect monocytes. To test this hypothesis, a mutant Vpx protein, called H82A, was constructed using mutagenic primers. This mutant lacks the ability to bind to APOBEC3A. Once created, the plasmid containing Vpx- H82A was transfected into 293T cells along with APOBEC3A. Vpx without the mutation, along with ABOBEC3A, was also expressed in 293T cells. Immunoblotting was utilized to visualize these results and determine if Vpx mediated degradation of APOBEC3A. We determined that neither wild type HIV-2 Vpx nor Vpx-H82A altered APOBEC3A levels. Currently, we are exploring the sensitivity of APOBEC3A to Vpx from other lentiviruses such as SIVmac, a simian immunodeficiency virus affecting Rhesus macaques. The same methods will be employed and an expression plasmid for the Vpx-H82A has been made. This project will contribute to our understanding of the innate immune response to lentiviral infection.
Effects of Cardiac Arrhythmic Mutant D95V on Calmodulin Structure and Stability
CSU Ballroom
APOBEC3A is a catalytically active DNA cytosine deaminase expressed in monocyte immune cells. This function allows APOBEC3A to mutate and restrict viruses, potentially including HIV. HIV-1, the causative agent of the major HIV/AIDS pandemic, is incapable of infecting monocytes. HIV-2, a less common variant, is capable of infecting monocytes. The unique protein Vpx, produced by HIV-2, but not HIV-1, is thought to be responsible for allowing HIV-2 infection in this immune cell. The objective of this study is to determine the sensitivity of human APOBEC3A to HIV-2 Vpx. We hypothesize that HIV-2 Vpx will be capable of mediating degradation of APOBEC3A and limiting its mutagenic capabilities, thus allowing HIV-2 to infect monocytes. To test this hypothesis, a mutant Vpx protein, called H82A, was constructed using mutagenic primers. This mutant lacks the ability to bind to APOBEC3A. Once created, the plasmid containing Vpx- H82A was transfected into 293T cells along with APOBEC3A. Vpx without the mutation, along with ABOBEC3A, was also expressed in 293T cells. Immunoblotting was utilized to visualize these results and determine if Vpx mediated degradation of APOBEC3A. We determined that neither wild type HIV-2 Vpx nor Vpx-H82A altered APOBEC3A levels. Currently, we are exploring the sensitivity of APOBEC3A to Vpx from other lentiviruses such as SIVmac, a simian immunodeficiency virus affecting Rhesus macaques. The same methods will be employed and an expression plasmid for the Vpx-H82A has been made. This project will contribute to our understanding of the innate immune response to lentiviral infection.
Recommended Citation
Rachuy, Jacob. "Effects of Cardiac Arrhythmic Mutant D95V on Calmodulin Structure and Stability." Undergraduate Research Symposium, Mankato, MN, April 11, 2017.
https://cornerstone.lib.mnsu.edu/urs/2017/poster-session-A/25
