Quantified expression of the ITPA enzyme mutations R178C within human neuron cells
Faculty Mentor
Dr. Nicholas Burgis
Presentation Type
Poster
Start Date
5-8-2024 9:00 AM
End Date
5-8-2024 10:45 AM
Location
PUB NCR
Primary Discipline of Presentation
Chemistry and Biochemistry
Abstract
Human inosine triphosphate pyrophosphatase, or ITPA, is an enzyme encoded by the ITPA gene, which plays a crucial role in nucleotide metabolism through the removal of abnormal purines. ITPA regulates cellular levels of the abnormal nucleotide inosine triphosphate (ITP) by hydrolyzation to the monophosphate form (IMP). Severe mutations in the ITPA gene result in the impairment of the ITPA enzyme, leading to the accumulation of ITP within the cell. The R178C mutation is a lethal variant, where cysteine replaces arginine at position 178, destabilizing the enzyme and rendering it inactive. These high levels of ITP can disrupt nucleotide, DNA, and RNA metabolism, ultimately culminating in early infantile encephalopathy, cardiac defects, and eventual mortality. Exposure to the noncanonical purine 6-N-hydroxylaminopurine (HAP) within cells is cytotoxic and a mutagen in addition to inhibiting DNA synthesis. While the damage resulting from the ITPA enzyme mutation is well observed, it is not understood how the defect is expressed within the cell. To better understand how this mutation causes infantile encephalopathy, we will use cell cultures exposed to HAP and MG132 (a proteasome inhibitor) to track cell viability and protein expression through fluorescent plate analysis, and mRNA expression levels of cell lysate using quantitative-Polymerase Chain Reaction (qPCR). The qPCR results will provide further direction to explore the mechanism and expression of ITPA gene mutations and improve understanding of the mechanisms of this enzyme opening doors to potential methods of treatment for affected individuals.
Recommended Citation
Kennerly, Sylvia H. and Kincaid, Rylee, "Quantified expression of the ITPA enzyme mutations R178C within human neuron cells" (2024). 2024 Symposium. 32.
https://dc.ewu.edu/srcw_2024/ps_2024/p1_2024/32
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Quantified expression of the ITPA enzyme mutations R178C within human neuron cells
PUB NCR
Human inosine triphosphate pyrophosphatase, or ITPA, is an enzyme encoded by the ITPA gene, which plays a crucial role in nucleotide metabolism through the removal of abnormal purines. ITPA regulates cellular levels of the abnormal nucleotide inosine triphosphate (ITP) by hydrolyzation to the monophosphate form (IMP). Severe mutations in the ITPA gene result in the impairment of the ITPA enzyme, leading to the accumulation of ITP within the cell. The R178C mutation is a lethal variant, where cysteine replaces arginine at position 178, destabilizing the enzyme and rendering it inactive. These high levels of ITP can disrupt nucleotide, DNA, and RNA metabolism, ultimately culminating in early infantile encephalopathy, cardiac defects, and eventual mortality. Exposure to the noncanonical purine 6-N-hydroxylaminopurine (HAP) within cells is cytotoxic and a mutagen in addition to inhibiting DNA synthesis. While the damage resulting from the ITPA enzyme mutation is well observed, it is not understood how the defect is expressed within the cell. To better understand how this mutation causes infantile encephalopathy, we will use cell cultures exposed to HAP and MG132 (a proteasome inhibitor) to track cell viability and protein expression through fluorescent plate analysis, and mRNA expression levels of cell lysate using quantitative-Polymerase Chain Reaction (qPCR). The qPCR results will provide further direction to explore the mechanism and expression of ITPA gene mutations and improve understanding of the mechanisms of this enzyme opening doors to potential methods of treatment for affected individuals.
