Treating Mutation E900X in Gene CUL4B using G-418
School Name
Governor's School for Science and Mathematics
Grade Level
12th Grade
Presentation Topic
Cell and Molecular Biology
Presentation Type
Mentored
Oral Presentation Award
2nd Place
Written Paper Award
3rd Place
Abstract
CUL4B is one of the most commonly mutated genes that results in x-linked intellectual disability. Mutations in CUL4B result in symptoms such as short stature, hypogonadism, abnormal gait, speech delay, prominent lower lip, and tremors. Because it is a single-base nucleotide mutation resulting in a premature stop codon, the approach to treatment is to attempt to create a “read through mutation”. Previous studies have tested treatments on diseases with premature stop codons and found that the “read through” effect can restore function to various different proteins. It has been shown to be affective in diseases such as cystic fibrosis, Duchenne muscular dystrophy, hemophilia, and cystinosis (Zingman et.al, 2007). The “read through” effect is when a protein continues to be created even in the presence of a premature stop codon. In this study, we used G-418, an aminoglycoside, to see if it can allow read-through of the stop codon. These ideas were tested using Western Blotting, which tested for the protein size, along with Immunoflourcence, which tested where the protein was located in the cell. The Western Blots confirmed that mutation E900X does in fact result in a truncated protein. The study also showed that CUL4B protein is usually localized in the nucleus for both the Wild Type Cells and the mutant cells. Interestingly, the E900X cells had a larger abundance than both the treated cells and the healthy cells. The next steps are to observe the effects of G-418 on protein size and to understand why the E900X cells proliferated.
Recommended Citation
Guest, Emelee, "Treating Mutation E900X in Gene CUL4B using G-418" (2018). South Carolina Junior Academy of Science. 19.
https://scholarexchange.furman.edu/scjas/2018/all/19
Location
Neville theater
Start Date
4-14-2018 12:15 PM
Presentation Format
Oral and Written
Treating Mutation E900X in Gene CUL4B using G-418
Neville theater
CUL4B is one of the most commonly mutated genes that results in x-linked intellectual disability. Mutations in CUL4B result in symptoms such as short stature, hypogonadism, abnormal gait, speech delay, prominent lower lip, and tremors. Because it is a single-base nucleotide mutation resulting in a premature stop codon, the approach to treatment is to attempt to create a “read through mutation”. Previous studies have tested treatments on diseases with premature stop codons and found that the “read through” effect can restore function to various different proteins. It has been shown to be affective in diseases such as cystic fibrosis, Duchenne muscular dystrophy, hemophilia, and cystinosis (Zingman et.al, 2007). The “read through” effect is when a protein continues to be created even in the presence of a premature stop codon. In this study, we used G-418, an aminoglycoside, to see if it can allow read-through of the stop codon. These ideas were tested using Western Blotting, which tested for the protein size, along with Immunoflourcence, which tested where the protein was located in the cell. The Western Blots confirmed that mutation E900X does in fact result in a truncated protein. The study also showed that CUL4B protein is usually localized in the nucleus for both the Wild Type Cells and the mutant cells. Interestingly, the E900X cells had a larger abundance than both the treated cells and the healthy cells. The next steps are to observe the effects of G-418 on protein size and to understand why the E900X cells proliferated.
