Mitochondria are subcellular compartments that are critical for energy production, cell signaling, and the biosynthesis of protein cofactors in higher eukaryotic cells. The mitochondrial DNA (mtDNA) genome is indispensable for mitochondrial function because it encodes protein subunits of the oxidative phosphorylation system and a full set of transfer and ribosomal RNAs. mtDNA degradation is an essential mechanism in mitochondrial genomic maintenance and a quality control measure to cope with mitochondrial DNA damage sourced from endogenous and environmental chemicals. The mechanism of mtDNA degradation and factors involved are poorly understood, which represents a significant knowledge gap. Such knowledge is fundamental to the understanding of mitochondrial genomic maintenance and pathology, because mtDNA degradation may contribute to the etiology of mtDNA depletion syndromes and to the activation of the innate immune system by circulating mtDNA. The objective of this project is to define the chemical and molecular basis of damaged mtDNA degradation and to clarify the role of a major transcription factor and DNA packaging protein TFAM (mitochondrial transcription factor A) in DNA degradation and repair. Addressing this critical knowledge gap will facilitate the PI?s long-term goal of unraveling the basis of mitochondrial DNA turnover and its role in mitochondrial pathobiology. This project focuses on a ubiquitous DNA lesion and central DNA repair intermediate, i.e. abasic (AP) sites. The central hypothesis is that TFAM modulates the stability of AP lesions and mediates AP-DNA degradation. This hypothesis is grounded in both strong preliminary data and empirical evidence. Preliminary results will be further evaluated by using a combination of quantitative biochemical, computational, and cellular approaches. Specifically, this research program will delineate the chemical and kinetic basis of TFAM-mediated AP-DNA destabilization, describe the involvement of TFAM in AP-DNA degradation in human cells, and clarify the regulatory role of TFAM in mtDNA repair. The expected outcome is that the project will fill a critical knowledge gap concerning the chemical and molecular mechanisms of mtDNA degradation and novel protein factors involved in the process. This application builds on the PI?s strong background in DNA and protein biochemistry, mechanistic enzymology, and quantitative analysis, and accelerates the progress in an exciting area of research into mitochondrial biology. The significance of this project is that it will define the chemical and molecular basis of an mtDNA-degradation pathway and the role of TFAM in mtDNA degradation and repair. Considering that AP sites are key intermediates in mtDNA repair, insights into AP-DNA degradation will have broad implications for understanding mitochondrial genomic maintenance and instability. New knowledge from this research will profoundly advance the field of mtDNA maintenance and potentially inform the development novel therapeutics for mitochondrial diseases.

Public Health Relevance

Mitochondrial dysfunction is known to cause more than 200 named disorders, and is associated with aging, neurodegeneration, cardiovascular disease, and diabetes mellitus. Mitochondrial DNA degradation is an important, but poorly understood process. The application seeks to decipher the chemical and molecular basis of mtDNA degradation, which is important for understanding mitochondrial genomic maintenance and developing new therapeutics for mitochondrial diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Unknown (R35)
Project #
3R35GM128854-03S1
Application #
10212125
Study Section
Special Emphasis Panel (ZGM1)
Program Officer
Ansong, Charles Kwaku
Project Start
2018-09-01
Project End
2023-08-31
Budget Start
2020-09-01
Budget End
2021-08-31
Support Year
3
Fiscal Year
2021
Total Cost
Indirect Cost
Name
University of California Riverside
Department
Chemistry
Type
Earth Sciences/Resources
DUNS #
627797426
City
Riverside
State
CA
Country
United States
Zip Code
92521