Event Title
Relationship of Vpx and APOBEC3A
Location
CSU 202
Start Date
18-4-2016 1:05 PM
End Date
18-4-2016 2:05 PM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Allison Land
Mentor's Department
Chemistry and Geology
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 solely by HIV-2, is thought to be responsible for allowing HIV-2 infection in this immune cell. I hypothesize that wild-type Vpx is capable of degrading APOBEC3A and limiting its mutagenic capabilities, thus allowing HIV-2 to infect monocytes. To test this hypothesis, a mutant Vpx protein, called H82A, is being constructed using mutagenic primers. This mutant lacks the ability to bind to APOBEC3A. Once created, the plasmid containing the mutated Vpx will be expressed in 293T cells along with APOBEC3A. Vpx without the mutation, along with ABOBEC3A, will also be expressed in 293T cells. Immunoblotting will be utilized to visualize these results and confirm the degradation, or lack of degradation of APOBEC3A. By comparing the expression levels of APOBEC3A in cells with H82A mutant Vpx to cells with wild type Vpx, it can be shown whether Vpx binding to APOBEC3A has an effect on degradation of APOBEC3A. This project will develop a model which will be used to assess the HIV-1 and HIV-2 restrictive abilities of APOBEC3A in human monocytes.
Relationship of Vpx and APOBEC3A
CSU 202
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 solely by HIV-2, is thought to be responsible for allowing HIV-2 infection in this immune cell. I hypothesize that wild-type Vpx is capable of degrading APOBEC3A and limiting its mutagenic capabilities, thus allowing HIV-2 to infect monocytes. To test this hypothesis, a mutant Vpx protein, called H82A, is being constructed using mutagenic primers. This mutant lacks the ability to bind to APOBEC3A. Once created, the plasmid containing the mutated Vpx will be expressed in 293T cells along with APOBEC3A. Vpx without the mutation, along with ABOBEC3A, will also be expressed in 293T cells. Immunoblotting will be utilized to visualize these results and confirm the degradation, or lack of degradation of APOBEC3A. By comparing the expression levels of APOBEC3A in cells with H82A mutant Vpx to cells with wild type Vpx, it can be shown whether Vpx binding to APOBEC3A has an effect on degradation of APOBEC3A. This project will develop a model which will be used to assess the HIV-1 and HIV-2 restrictive abilities of APOBEC3A in human monocytes.
Recommended Citation
Rachuy, Jacob. "Relationship of Vpx and APOBEC3A." Undergraduate Research Symposium, Mankato, MN, April 18, 2016.
https://cornerstone.lib.mnsu.edu/urs/2016/oral-session-09/1
