Mitochondria are required for a large number of cellular functions, including ion homeostasis, respiration, and programmed cell death. Consequently, defects in mitochondrial function have emerged as causative or contributing factors in a growing number of diverse human diseases such as cancer, cardiomyopathies, metabolic syndrome, and various neurodegenerative disorders. These diseases affect more than 50 million adults in the United States. Ad hoc proteomic and genetic studies in mammalian system have uncovered new mitochondrial protein (MP) complexes or pathways, but yet there has been no systematic experimental study of the mitochondrial interactome in mammalian system describing how these proteins function together in networks of pathways and complexes. Furthermore, it is also difficult to pinpoint the role of mitochondrial dysfunction in human disease because the interactions of MPs, both within and outside the mitochondria, are extensive and can be difficult to detect. This proposal seeks to begin addressing this deficit by creating a detailed physical (protein-protein) and genetic (gene-gene) interaction maps among MPs, which will help determine how mitochondrial protein complexes, within the framework of higher order networks, regulate and execute the associated processes. Using an optimized lentivirus-delivered tagging system coupled with mass spectrometry, a mammalian mitochondrial physical interactome map will be created using both native and mutant MPs with known association with human diseases. The resulting interaction networks will be compared to identify protein candidates that are relevant to disease onset and progression, and assess for variation in putative posttranslational modification sites involved in the progression of mitochondrial diseases (Aim1). Because genetic interaction (GI) is critical for revealing pathway-level relationships, optimized high precision quantitative pooled shRNAi coupled with deep sequencing approach, pioneered by the Weissman laboratory, will be used to query genes encoding for MP function by comparing the growth of pooled shRNAi treated cells on glucose relative to galactose as carbon sources in the media (Aim 2). This GI screening procedure will uncover new candidates that can toggle the glycolysis/mitochondrial respiration switch, which can be harnessed for therapeutic intervention. Finally, the GI data resulting from Aim 2 will be integrated with the proteomics data from Aim 1 (Aim 3) to investigate the functional relatedness and overall pathway architecture of the MP complexes to understand the fundamental mitochondrial biology and the role of mitochondrial dysfunction in disease. Collectively, these objectives are designed to provide new insights into the complex etiologies of diseases and have the potential to identify novel therapeutic avenues.

Public Health Relevance

Mitochondria are subcellular organelles that perform a myriad of diverse, essential functions in cells. Thus, it is not surprising that mitochondrial dysfunctionis emerging as a causative factor in a wide range of human diseases, including common diseases, such as cancer, metabolic, and neurodegeneration. The proposed work will address how mitochondrial dysfunction causes human disease by exploring the systems properties of the mitochondrial organelle and is relevant to the part of the NIH's mission that fosters fundamental basic cell biology discoveries that will directly lead to the identification of new therapeutic tarets and therapies for the treatment of wide array of human diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM106019-01
Application #
8490217
Study Section
Special Emphasis Panel (ZGM1-CBB-0 (MI))
Program Officer
Anderson, Vernon
Project Start
2013-09-20
Project End
2017-05-31
Budget Start
2013-09-20
Budget End
2014-05-31
Support Year
1
Fiscal Year
2013
Total Cost
$240,629
Indirect Cost
$65,629
Name
University of California Davis
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618
Friedman, Jonathan R; Nunnari, Jodi (2014) Mitochondrial form and function. Nature 505:335-43
Labbé, Katherine; Murley, Andrew; Nunnari, Jodi (2014) Determinants and functions of mitochondrial behavior. Annu Rev Cell Dev Biol 30:357-91