Technical knockout, a Drosophila model of mitochondrial deafness.

Mutations in mtDNA-encoded components of the mitochondrial translational apparatus are associated with diverse pathological states in humans, notably sensorineural deafness. To develop animal models of such disorders, we have manipulated the nuclear gene for mitochondrial ribosomal protein S12 in Drosophila (technical knockout, tko). The prototypic mutant tko(25t) exhibits developmental delay, bang sensitivity, impaired male courtship, and defective response to sound. On the basis of a transgenic reversion test, these phenotypes are attributable to a single substitution (L85H) at a conserved residue of the tko protein. The mutant is hypersensitive to doxycyclin, an antibiotic that selectively inhibits mitochondrial protein synthesis, and mutant larvae have greatly diminished activities of mitochondrial redox enzymes and decreased levels of mitochondrial small-subunit rRNA. A second mutation in the tko gene, Q116K, which is predicted to impair the accuracy of mitochondrial translation, results in the completely different phenotype of recessive female sterility, based on three independent transgenic insertions. We infer that the tko(25t) mutant provides a model of mitochondrial hearing impairment resulting from a quantitative deficiency of mitochondrial translational capacity.

[1]  P. Garland,et al.  [2] Citrate synthase from rat liver: [EC 4.1.3.7 Citrate oxaloacetage-lyase (CoA-acetylating)] , 1969 .

[2]  T. Kaufman,et al.  The anatomy and function of a segment of the X chromosome of Drosophila melanogaster. , 1972, Genetics.

[3]  T. Kaufman,et al.  Lethality patterns and morphology of selected lethal and semi-lethal mutations in the zeste-white region of Drosophila melanogaster. , 1972, Genetics.

[4]  L. Søndergaard,et al.  The effect of the Out‐coldtS mutation on temperature induced changes in the arrhenius activation energy of succinate‐cyto‐ chrome c reductase activity in drosophila , 1975, FEBS letters.

[5]  P. Dennis,et al.  Regulation of ribosomal protein synthesis in Escherichia coli B/r , 1975, Journal of bacteriology.

[6]  F. Schilcher,et al.  The role of auditory stimuli in the courtship of Drosophila melanogaster , 1976, Animal Behaviour.

[7]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[8]  J. Rubenstein,et al.  The mitochondrial DNA of drosophila melanogaster exists in two distinct and stable superhelical forms , 1977, Cell.

[9]  B. Errede,et al.  Preparation and properties of complex IV (ferrocytochrome c: oxygen oxidoreductase EC 1.9.3.1). , 1978, Methods in enzymology.

[10]  Y. Hatefi Preparation and properties of dihydroubiquinone: cytochrome c oxidoreductase (complex III). , 1978, Methods in enzymology.

[11]  Y. Hatefi Preparation and properties of NADH: ubiquinone oxidoreductase (complexI), EC 1.6.5.3. , 1978, Methods in enzymology.

[12]  S. Alziari,et al.  In vitro incorporation of (35S)-methionine in mitochondrial proteins of Drosophila melanogaster. , 1981, Biochemical and biophysical research communications.

[13]  B. Ganetzky,et al.  Indirect Suppression Involving Behavioral Mutants with Altered Nerve Excitability in DROSOPHILA MELANOGASTER. , 1982, Genetics.

[14]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[15]  M. Kelley,et al.  Mutagenesis in Oocytes of DROSOPHILA MELANOGASTER. I. Scheduled Synthesis of Nuclear and Mitochondrial DNA and Unscheduled DNA Synthesis. , 1983, Genetics.

[16]  M. Rosbash,et al.  A Drosophila Minute gene encodes a ribosomal protein , 1985, Nature.

[17]  Constance S. Royden,et al.  The tko locus, site of a behavioral mutation in D. melanogaster, codes for a protein homologous to prokaryotic ribosomal protein S12 , 1987, Cell.

[18]  C. Haanen,et al.  Mitochondrial biogenesis and mitochondrial activity during the progression of the cell cycle of human leukemic cells. , 1988, Experimental cell research.

[19]  B. Burnet,et al.  The effects on locomotor activity and reactivity of the hypoactive and inactive mutations of Drosophila melanogaster , 1988, Heredity.

[20]  J. Jallon,et al.  Effects of the hypoactive and inactive mutations on mating success in Drosophila melanogaster , 1989, Heredity.

[21]  S. Tourmente,et al.  Expression of six mitochondrial genes during Drosophila oogenesis: analysis by in situ hybridization. , 1990, Cell differentiation and development : the official journal of the International Society of Developmental Biologists.

[22]  A. Lundin,et al.  A luminometric method for the determination of ATP and phosphocreatine in single human skeletal muscle fibres. , 1991, Journal of bioluminescence and chemiluminescence.

[23]  D. Lindsley,et al.  The Genome of Drosophila Melanogaster , 1992 .

[24]  D. Wolstenholme,et al.  Animal mitochondrial DNA: structure and evolution. , 1992, International review of cytology.

[25]  Mordechai Shohat,et al.  Mitochondrial ribosomal RNA mutation associated with both antibiotic–induced and non–syndromic deafness , 1993, Nature Genetics.

[26]  J. Palka,et al.  A mutation of the drosophila sodium pump α subunit gene results in bang-sensitive paralysis , 1994, Neuron.

[27]  M. Tanouye,et al.  The Drosophila easily shocked gene: A mutation in a phospholipid synthetic pathway causes seizure, neuronal failure, and paralysis , 1994, Cell.

[28]  M Heisenberg,et al.  THE CENTRAL COMPLEX OF DROSOPHILA MELANOGASTER IS INVOLVED IN FLIGHT CONTROL: STUDIES ON MUTANTS AND MOSAICS OF THE GENE ELLIPSOID BODY OPEN , 1994, Journal of neurogenetics.

[29]  J. C. Hall The mating of a fly. , 1994, Science.

[30]  Winfried Denk,et al.  Calcium imaging of single stereocilia in hair cells: Localization of transduction channels at both ends of tip links , 1995, Neuron.

[31]  S. McRobert,et al.  Selection for increased female sexual receptivity inRaised stocks ofDrosophila melanogaster , 1995, Behavior genetics.

[32]  M. Tomaru,et al.  Heterospecific inter-pulse intervals of courtship song elicit female rejection in Drosophila biauraria , 1995, Animal Behaviour.

[33]  M. Tanouye,et al.  Seizures and failures in the giant fiber pathway of Drosophila bang- sensitive paralytic mutants , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[34]  C. Rushlow,et al.  The transcriptional corepressor DSP1 inhibits activated transcription by disrupting TFIIA‐TBP complex formation. , 1996, The EMBO journal.

[35]  D. Johns,et al.  Clinical Spectrum of Mitochondrial Diseases , 1996, Seminars in neurology.

[36]  N. Perrimon,et al.  A genetic screen for mutations that disrupt an auditory response in Drosophila melanogaster. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Nektarios Tavernarakis,et al.  Molecular modeling of mechanotransduction in the nematode Caenorhabditis elegans. , 1997, Annual review of physiology.

[38]  J. Mazat,et al.  Mitochondria Are Excitable Organelles Capable of Generating and Conveying Electrical and Calcium Signals , 1997, Cell.

[39]  Z. Shah,et al.  Metazoan nuclear genes for mitoribosomal protein S12. , 1997, Gene.

[40]  S. Saebøe-Larssen,et al.  Ribosomal protein insufficiency and the minute syndrome in Drosophila: a dose-response relationship. , 1998, Genetics.

[41]  J. Mazat,et al.  Modulation of cell calcium signals by mitochondria. , 1998 .

[42]  K. Majamaa,et al.  Epidemiology of A3243G, the mutation for mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes: prevalence of the mutation in an adult population. , 1998, American journal of human genetics.

[43]  K. Steel,et al.  The molecular genetics of inherited deafness – current knowledge and recent advances , 1998, The Journal of Laryngology & Otology.

[44]  J. Toivonen,et al.  Modelling in Escherichia coli of mutations in mitoribosomal protein S12: novel mutant phenotypes of rpsL , 1999, Molecular microbiology.

[45]  D. Eberl,et al.  Feeling the vibes: chordotonal mechanisms in insect hearing , 1999, Current Opinion in Neurobiology.

[46]  N. Fischel‐Ghodsian Mitochondrial deafness mutations reviewed , 1999, Human mutation.

[47]  M. Ritchie,et al.  Drosophila song as a species-specific mating signal and the behavioural importance of Kyriacou & Hall cycles in D.melanogaster song , 1999, Animal Behaviour.

[48]  J. Toivonen,et al.  Expression of the Gene for Mitoribosomal Protein S12 Is Controlled in Human Cells at the Levels of Transcription, RNA Splicing, and Translation* , 1999, The Journal of Biological Chemistry.

[49]  C. Zuker,et al.  A Drosophila mechanosensory transduction channel. , 2000, Science.