G418 Treatment for The Suppression of a Nonsense Mutation in the X-Linked Intellectual Disability Gene CUL4B

School Name

Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Physiology and Health

Presentation Type

Mentored

Mentor

Mentor: Anand Srivastava, Greenwood Genetic Center

Abstract

Intellectual disabilities affect approximately 1-3% of the global population, and nonsense mutations in various regions of the X chromosome are primary causes of these diseases. Nonsense mutations are changes to DNA that create premature stop codons, which code for truncated proteins that are often non-functional. Studies have shown that people with a nonsense mutation in the CUL4B gene, which normally encodes a cullin protein and promotes the ubiquitination of several histones, have a syndromic intellectual disability. Some of the symptoms associated with the mutant phenotype are enlarged head, abnormal gait, and short stature. One proposed treatment approach for genetic diseases like syndromic intellectual disability is to suppress the nonsense mutation causing the disease by administering a low molecular weight drug. Previous research has shown that aminoglycosides bind to the ribosome and allow protein elongation to continue even in the presence of a premature stop codon. Despite this discovery, aminoglycosides have not been widely-tested to combat intellectual disability. We examined the ability of the aminoglycoside, G418, to treat kidney cells that contained a nonsense mutation in CUL4B. Western Blots showed that G418 successfully produced a read-through of CUL4B wild-type protein from a mutant CUL4B gene, and immunofluorescence determined that the nonsense mutation did not affect the normal localization of CUL4B. Further research can examine the ability of G418 to produce read-through from other nonsense mutations. Additionally, future research can investigate how to minimize the side effects from aminoglycoside toxicity.

Location

Wall 318

Start Date

3-25-2017 10:00 AM

Presentation Format

Oral and Written

Group Project

No

COinS
 
Mar 25th, 10:00 AM

G418 Treatment for The Suppression of a Nonsense Mutation in the X-Linked Intellectual Disability Gene CUL4B

Wall 318

Intellectual disabilities affect approximately 1-3% of the global population, and nonsense mutations in various regions of the X chromosome are primary causes of these diseases. Nonsense mutations are changes to DNA that create premature stop codons, which code for truncated proteins that are often non-functional. Studies have shown that people with a nonsense mutation in the CUL4B gene, which normally encodes a cullin protein and promotes the ubiquitination of several histones, have a syndromic intellectual disability. Some of the symptoms associated with the mutant phenotype are enlarged head, abnormal gait, and short stature. One proposed treatment approach for genetic diseases like syndromic intellectual disability is to suppress the nonsense mutation causing the disease by administering a low molecular weight drug. Previous research has shown that aminoglycosides bind to the ribosome and allow protein elongation to continue even in the presence of a premature stop codon. Despite this discovery, aminoglycosides have not been widely-tested to combat intellectual disability. We examined the ability of the aminoglycoside, G418, to treat kidney cells that contained a nonsense mutation in CUL4B. Western Blots showed that G418 successfully produced a read-through of CUL4B wild-type protein from a mutant CUL4B gene, and immunofluorescence determined that the nonsense mutation did not affect the normal localization of CUL4B. Further research can examine the ability of G418 to produce read-through from other nonsense mutations. Additionally, future research can investigate how to minimize the side effects from aminoglycoside toxicity.