Depletion of mitochondrial DNA alters glucose metabolism in SK-Hep1 cells.

Maternally inherited mitochondrial DNA (mtDNA) has been suggested to be a genetic factor for diabetes. Reports have shown a decrease of mtDNA content in tissues of diabetic patients. We investigated the effects of mtDNA depletion on glucose metabolism by use of rho(0) SK-Hep1 human hepatoma cells, whose mtDNA was depleted by long-term exposure to ethidium bromide. The rho(0) cells failed to hyperpolarize mitochondrial membrane potential in response to glucose stimulation. Intracellular ATP content, glucose-stimulated ATP production, glucose uptake, steady-state mRNA and protein levels of glucose transporters, and cellular activities of glucose-metabolizing enzymes were decreased in rho(0) cells compared with parental rho(+) cells. Our results suggest that the quantitative reduction of mtDNA may suppress the expression of nuclear DNA-encoded glucose transporters and enzymes of glucose metabolism. Thus this may lead to diabetic status, such as decreased ATP production and glucose utilization.

[1]  R. N. Bergman,et al.  Role of glucose and insulin resistance in development of type 2 diabetes mellitus: results of a 25-year follow-up study , 1992, The Lancet.

[2]  D. Brdiczka.  Function of the outer mitochondrial compartment in regulation of energy metabolism. , 1994, Biochimica et biophysica acta.

[3]  S. Penman,et al.  Selective inhibition of the synthesis of mitochondria-associated RNA by ethidium bromide. , 1969, Journal of molecular biology.

[4]  J. Hayashi,et al.  Mitochondrial DNA Is Required for Regulation of Glucose-stimulated Insulin Secretion in a Mouse Pancreatic Beta Cell Line, MIN6* , 1996, The Journal of Biological Chemistry.

[5]  R. Perham,et al.  Glyceraldehyde 3-phosphate dehydrogenases: I. The protein chains in glyceraldehyde 3-phosphate dehydrogenase from pig muscle , 1965 .

[6]  R. Grobholz,et al.  Reduction in the expression of glucose transporter protein GLUT 2 in preneoplastic and neoplastic hepatic lesions and reexpression of GLUT 1 in late stages of hepatocarcinogenesis. , 1993, Cancer research.

[7]  C. Godinot,et al.  Functional F1-ATPase essential in maintaining growth and membrane potential of human mitochondrial DNA-depleted rho degrees cells. , 1998, The Journal of biological chemistry.

[8]  C. Wollheim,et al.  Effects of depletion of mitochondrial DNA in metabolism secretion coupling in INS-1 cells. , 1998, Diabetes.

[9]  Y. Yazaki,et al.  Ethidium Bromide-induced Inhibition of Mitochondrial Gene Transcription Suppresses Glucose-stimulated Insulin Release in the Mouse Pancreatic β-Cell Line βHC9* , 1998, The Journal of Biological Chemistry.

[10]  C. Godinot,et al.  Functional F1-ATPase Essential in Maintaining Growth and Membrane Potential of Human Mitochondrial DNA-depleted ρ° Cells* , 1998, The Journal of Biological Chemistry.

[11]  John Eric Wilson,et al.  Further studies on the coupling of mitochondrially bound hexokinase to intramitochondrially compartmented ATP, generated by oxidative phosphorylation. , 1998, Archives of biochemistry and biophysics.

[12]  B. Storrie,et al.  The preparative isolation of mitochondria from Chinese hamster ovary cells. , 1987, Analytical biochemistry.

[13]  R. Vaughan-Jones,et al.  Effects of mitochondrial uncouplers on intracellular calcium, pH and membrane potential in rat carotid body type I cells , 1998, The Journal of physiology.

[14]  M. King,et al.  Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. , 1989, Science.

[15]  M. Jensen,et al.  Effects of type 2 diabetes on the ability of insulin and glucose to regulate splanchnic and muscle glucose metabolism: evidence for a defect in hepatic glucokinase activity. , 2000, Diabetes.

[16]  P. Owen,et al.  Factors influencing the activity of succinate dehydrogenase in membrane preparations from Micrococcus lysodeikticus. , 1970, The Biochemical journal.

[17]  B. Kahn.  Glucose Transport: Pivotal Step in Insulin Action , 1996, Diabetes.

[18]  D. Galton,et al.  ASSOCIATION OF GENETIC VARIANT OF THE GLUCOSE TRANSPORTER WITH NON-INSULIN-DEPENDENT DIABETES MELLITUS , 1988, The Lancet.

[19]  Y. Mita,et al.  Utility of 3-O-methyl-N-acetyl-D-glucosamine, an N-acetylglucosamine kinase inhibitor, for accurate assay of glucokinase in pancreatic islets and liver. , 1994, Enzyme & protein.

[20]  S. Dimauro,et al.  mtDNA depletion with variable tissue expression: a novel genetic abnormality in mitochondrial diseases. , 1991, American journal of human genetics.

[21]  R. D. Demoss.  [43] Glucose-6-phosphate and 6-phosphogluconic dehydrogenases from Leuconostoc mesenteroides , 1955 .

[22]  K. Gempel,et al.  Mitochondria and Diabetes: Genetic, Biochemical, and Clinical Implications of the Cellular Energy Circuit , 1996, Diabetes.

[23]  N. Welsh,et al.  Mitochondrial deoxyribonucleic acid content is specifically decreased in adult, but not fetal, pancreatic islets of the Goto-Kakizaki rat, a genetic model of noninsulin-dependent diabetes. , 1995, Endocrinology.

[24]  D. Wallace,et al.  Mitochondrial diabetes revisited , 1994, Nature Genetics.

[25]  D. Wallace.  Mitochondrial genetics: a paradigm for aging and degenerative diseases? , 1992, Science.

[26]  C. Kahn,et al.  Increased expression of mitochondrial-encoded genes in skeletal muscle of humans with diabetes mellitus. , 1995, The Journal of clinical investigation.

[27]  K. Stuhlmeier,et al.  Importance of Glucose-6-phosphate Dehydrogenase Activity for Cell Growth* , 1998, The Journal of Biological Chemistry.

[28]  D. Wallace,et al.  Maternally transmitted diabetes and deafness associated with a 10.4 kb mitochondrial DNA deletion , 1992, Nature Genetics.

[29]  A. Wolf,et al.  Metabolic trapping as a principle of oradiopharmaceutical design: some factors resposible for the biodistribution of [18F] 2-deoxy-2-fluoro-D-glucose. , 1978, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[30]  M. Welsh,et al.  Exposure of pancreatic islets to different alkylating agents decreases mitochondrial DNA content but only streptozotocin induces long-lasting functional impairment of B-cells. , 1991, Biochemical pharmacology.

[31]  K. Gerbitz,et al.  Mitochondrial diabetes mellitus: a review. , 1995, Biochimica et biophysica acta.

[32]  M. Matsuhisa,et al.  Increased hepatic glucose production and decreased hepatic glucose uptake at the prediabetic phase in the Otsuka Long-Evans Tokushima fatty rat model. , 1998, Metabolism: clinical and experimental.

[33]  A. Klip,et al.  Mechanisms of adaptation of glucose transporters to changes in the oxidative chain of muscle and fat cells. , 1993, The American journal of physiology.

[34]  O. H. Bing,et al.  Lactate dehydrogenase and isoenzyme changes in rats with experimental thiamine deficiency. , 1976, Metabolism: clinical and experimental.

[35]  K. Park,et al.  Decreased mitochondrial DNA content in peripheral blood precedes the development of non-insulin-dependent diabetes mellitus. , 1998, Diabetes research and clinical practice.

[36]  R. Sakuta,et al.  A subtype of diabetes mellitus associated with a mutation of mitochondrial DNA. , 1994, The New England journal of medicine.