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Effect of a Deletion of Mitochondrial DNA on Cell Growth, Mitochondrial DNA Content, and Metabolic Activity
Author(s) -
Keck Christopher,
Gulfo Michelle K.,
Pogozelski Wendy
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.536.17
Subject(s) - mitochondrial dna , oxidative phosphorylation , mitochondrion , biology , glycolysis , metabolism , dna , cell culture , microbiology and biotechnology , cell , biochemistry , gene , genetics
Objective To determine the effects of human mitochondrial DNA (mtDNA) deletions such as the 4977‐bp “common deletion” (ΔmtDNA 4977 ) on cell growth, mtDNA content and metabolismIntroduction Human mitochondria (mt) contain circular DNA molecules of 16549 base pairs and can contain large deletions that correlate with aging and disease. We have studied the effects of a particular deletion of 4977‐bp (ΔmtDNA 4977 ),. The deleted mtDNA is missing regions of Complexes I, IV and V. We have been studying how this deletion affects cell growth, mtDNA content, and metabolism in a lymphoblast cell line bearing elevated levels of the deletion, comparing the effects with normal cells. Methods We used real‐time PCR to measure the amount of mtDNA bearing the deletion as well as total mtDNA in the two cell lines. We compared the growth and metabolism of the two cells lines by varying glucose content in media and measuring numbers of cells as a function of time. We measured the rates of glycolysis with a lactate assay, measured oxidative phosphorylation with an oxygen consumption assay, and studied the shapes of the cells via microscopy. We also investigated the effect of succinic acid on metabolism. Results Cells with elevated ΔmtDNA 4977 actually had more total mtDNA content than did normal cells, perhaps as a compensatory response. Despite this increased mtDNA content, they grew more slowly, grew poorly in media with low glucose, showed higher rates of glycolysis, formed spheroids more readily; and were more often polarized in shape. Normal control cells grew well in all media types tested; however, when grown in high glucose, normal control cells were polarized more often. Succinic acid supplementation did not substantially affect growth of either cell type. Initial results suggest that accumulation of this deletion increases the dependence of the cell on glucose metabolism and contributes to activation of mechanisms that result in projections and polarization. Support or Funding Information The Geneseo Foundation and Geneseo Student Association This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .