1887

Abstract

Summary: Aminoglycoside antibiotics, used to treat bacterial infections by interfering with proofreading during protein synthesis, cause sensorineural hearing loss in genetically susceptible individuals. The only aminoglycoside-hypersensitivity mutations which have been described in humans are in the mitochondrial 12S rRNA gene, potentially allowing increased antibiotic binding to mitochondrial ribosomes. To identify additional predisposing mutations, a yeast model system was used to isolate genes which interact with or bypass the effects of aminoglycoside antibiotics. A novel yeast gene was isolated which, in high copy, confers neomycin resistance to yeast transformants. The neomycin-resistance 1 gene () encodes a potential 1151 aa integral membrane protein, most homologous to the yeast gene product, a Ca-ATPase involved in cytoplasmic ribosome assembly. The N-terminus of Neo1p is partially homologous to abrin A-chain, another protein which interacts with cytoplasmic ribosomes. Mutagenesis experiments demonstrate that the product is essential for vegetative growth and that the drug-resistance phenotype requires ATPase function.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-12-3407
1996-12-01
2021-10-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/12/mic-142-12-3407.html?itemId=/content/journal/micro/10.1099/13500872-142-12-3407&mimeType=html&fmt=ahah

References

  1. von Ahsen U., Noller H. F. 1993; Footprinting the sites of interaction of antibiotics with catalytic group I intron RNA. Science 260:1500–1503
    [Google Scholar]
  2. von Ahsen U., Davies J., Schroeder R. 1991; Antibiotic inhibition of group I ribozyme function. Nature 353:368–370
    [Google Scholar]
  3. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410
    [Google Scholar]
  4. Bairoch A. 1991; PROSITE: a dictionary of sites and patterns in proteins. Nucleic Acids Res 19: suppl. 2241–2245
    [Google Scholar]
  5. Bairoch A., Boeckmann B. 1991; The SWISS-PROT protein sequence data bank. Nucleic Acids Res 19: suppl. 2247–2249
    [Google Scholar]
  6. Bacino C., Prezant T. R., Bu X., Fournier P., Fischel-Ghodsian N. 1995; Susceptibility mutations in the mitochondrial small ribosomal RNA gene in aminoglycoside induced deafness. Pharmacogenetics165–172
    [Google Scholar]
  7. Balzi E., Wang M., Leterme S., VanDyck L., Goffeau A. 1994; PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1. . J Biol Chem 269:2206–2214
    [Google Scholar]
  8. Bissinger P. H., Kuchler K. 1994; Molecular cloning and expression of the Saccharomyces cerevisiae STS 1 gene product: a yeast ABC transporter conferring mycotoxin resistance. J Biol Chem 269:4180–4186
    [Google Scholar]
  9. Botstein D., Falco S. G., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. 1979; Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–24
    [Google Scholar]
  10. Carlson M., Botstein D. 1982; Two differentially regulated mRNAs with different 5’ ends encode secreted with intracellular forms of yeast invertase. Cell 28:145–154
    [Google Scholar]
  11. Chaddock J. A., Lord J. M., Hartley M. R., Roberts L. M. 1994; Pokeweed antiviral protein (PAP) mutations which permit E. coli growth do not eliminate catalytic activity towards prokaryotic ribosomes. Nucleic Acids Res 22:1536–1540
    [Google Scholar]
  12. Chernoff Y. O., Vincent A., Liebman S. W. 1994; Mutations in eukaryotic 18S ribosomal RNA affect translational fidelity and resistance to aminoglycoside antibiotics. EMBO J 13:906–913
    [Google Scholar]
  13. Dayhoff M. O., Schwartz R. M., Orcutt B. C. 1978; A model of evolutionary change in proteins. In Atlas of Protein Sequence and Structure 5 suppl. 3 pp. 89–99 Dayhoff M. O. Edited by Washington, DC: National Biomedical Research Foundation.;
    [Google Scholar]
  14. Dean N. 1995; Yeast glycosylation mutants are sensitive to aminoglycosides. Proc Natl Acad Sci USA 92:1287–1291
    [Google Scholar]
  15. Endo Y., Mitsui K., Motizuki M., Tsurugi K. 1987; The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. The site and the characteristics of the modification in 28S ribosomal RNA caused by the toxins. J Biol Chem 262:5908–5912
    [Google Scholar]
  16. Evensen G., Mathiesen A. 1991; Direct molecular cloning and expression of two distinct abrin A-chains. J Biol Chem 266:6848–6852
    [Google Scholar]
  17. Garetz S. L., Rhee D. J., Schacht J. 1994; Sulfhydryl com¬ pounds and antioxidants inhibit cytotoxicity to outer hair cells of a gentamicin metabolite in vitro. . Hearing Res 77:75–80
    [Google Scholar]
  18. Higashi K. 1989; Unique inheritance of streptomycin-induced deafness. Clin Genet 35:433–436
    [Google Scholar]
  19. Hornig H., Woolley P., Luhrman R. 1987; Decoding at the ribosomal A site: antibiotics, misreading and energy of aminoacyl- tRNA binding. Biochimie 69:803–818
    [Google Scholar]
  20. Hu D. -N., Qiu W. -Q., Wu B. -T., Fang L. -Z., Zhou F., Gu Y. -P., Zhang Q. -H., Yan J. -H., Ding Y. -Q., Wong H. 1991; Genetic aspects of antibiotic induced deafness: mitochondrial inheritance. J Med Genet 28:79–83
    [Google Scholar]
  21. Hung C. -H., Lee M.-C, Lin J. -Y. 1993; Primary structure of three distinct isoabrins determined by cDNA sequencing. Conservation and significance. J Mol Biol 229:263–267
    [Google Scholar]
  22. Hutchin T., Haworth I., Higashi K., Fischel-Ghodsian N., Stoneking M., Saha N., Arnos C., Cortopassi G. 1993; A molecular basis for human hypersensitivity to aminoglycoside antibiotics. Nucleic Acids Res 21:4174–4179
    [Google Scholar]
  23. Ito H., Fukuda Y., Murata K., Kimura A. 1983; T ransformation of intact yeast cells treated with alkali cations. J Bacterial 153:163–168
    [Google Scholar]
  24. Kim Y., Robertus J. D. 1992; Analysis of several key active site residues of ricin A chain by mutagenesis and X-ray crystallography. Protein Eng 5:775–779
    [Google Scholar]
  25. Kyte J., Doolittle R. F. 1982; A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
    [Google Scholar]
  26. Leppert G., McDevitt R., Falco S. C., Ficke M. B., Golin J. 1990; Cloning by gene amplification of two loci conferring multiple drug resistance in Saccharomyces. . Genetics 125:13–20
    [Google Scholar]
  27. Li M., Tzagoloff A., Underbrink-Lyon K., Martin N. C. 1982; Identification of the paromomycin-resistance mutation in the 15S rRNA gene of yeast mitochondria. J Biol Chem 257:5921–5928
    [Google Scholar]
  28. McCusker J. H., Perlin D. S., Haber J. E. 1987; Pleiotropic plasma membrane ATPase mutations of Saccharomyces cerevisiae. . Mol Cell Biol 7:4082–4088
    [Google Scholar]
  29. Marbois B. N., Hsu A., Pillai R., Colicelli J., Clarke C. F. 1994; Cloning of a rat cDNA encoding dihydroxypolyprenylbenzoate methyltransferase by functional complementation of a Saccharomyces cerevisiae mutant deficient in ubiquinone biosynthesis. Gene 138:213–217
    [Google Scholar]
  30. Moazed D., Noller H. F. 1987; Interaction of antibiotics with functional sites in 16S ribosomal RNA. Nature 327:389–394
    [Google Scholar]
  31. Monzingo A. F., Robertus J. D. 1992; X-ray analysis of substrate analogs in the ricin A-chain active site. J Mol Biol 227:1136–1145
    [Google Scholar]
  32. Monzingo A. F., Collins E. J., Ernst S. R., Irvin J. D., Robertus J. D. 1993; The 2-5 A structure of pokeweed antiviral protein. J Mol Biol 233:705–715
    [Google Scholar]
  33. Nakai K., Kanehisa M. 1992; A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics 14:897–911
    [Google Scholar]
  34. Pearson W. R., Lipman D. J. 1988; Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
    [Google Scholar]
  35. Pedersen P. L. 1987; Ion motive ATPases. I. Ubiquity, properties, and significance to cell function. Trends Biochem Sci 12:146–150
    [Google Scholar]
  36. Perlin D. S., Brown C. L., Haber J. E. 1988; Membrane potential defect in hygromycin B-resistant pma1 mutants of Saccharomyces cerevisiae. . J Biol Chem 263:18118–18122
    [Google Scholar]
  37. Perlin D. S., Harris S. L., Seto-Young D., Haber J. E. 1989; Defective H( + )-ATPase of hygromycin B-resistant pma1 mutants from Saccharomyces cerevisiae. . J Biol Chem 264:21857–21864
    [Google Scholar]
  38. Prezant T. R., Agapian J. V., Bohlman M. C. 1993; Mitochondrial ribosomal RNA mutation associated with both antibiotic-induced and non-syndromic deafness. Nature Genet 4:289–294
    [Google Scholar]
  39. Ready M. P., Kim Y., Robertus J. D. 1991; Site-directed mutagenesis of ricin A-chain and implications for the mechanism of action. Proteins 10:270–278
    [Google Scholar]
  40. Ripmaster T. L., Vaughn G. P., Woolford J. L. 1993; DRS1 to DRS7, novel genes required for ribosome assembly and function in Saccharomyces cerevisiae. . Mol Cell Biol 13:7901–7912
    [Google Scholar]
  41. Rothstein R. 1991; Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol 194:281–301
    [Google Scholar]
  42. Rudolph H. K., Antebi A., Fink G. R., Buckley C. M., Dorman T. E., LeVitre J., Davidow L. S., Mao J. -l., Moir D. T. 1989; The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family. Cell 58:133–145
    [Google Scholar]
  43. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B. 1988; Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  44. Sambrook J., Fritsch E. F. 1989 Molecular Cloning: a Eahoratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  45. Sandbaken M. G., Lupisella J. A., DiDomenico B., Chakraburtty K. 1990; Protein synthesis in yeast. Structural and functional analysis of the gene encoding elongation factor 3. J Biol Chem 265:15838–15844
    [Google Scholar]
  46. Sande M. A., Mandell G. L. 1990; Antimicrobial agents. In The Pharmacological Basis of Therapeutics,, 8th edn. pp. 1098–1116 Gilman A. G., Rail T. W., Nies A. S., Taylor P. Edited by Elmsford, NY: Pergamon Press.;
    [Google Scholar]
  47. Schacht J., Weiner N. 1986; Aminoglycoside-induced hearing loss: a molecular hypothesis. J Oto-Rhino-Laryngol & Related Specialties 48:116–123
    [Google Scholar]
  48. Serrano R. 1991; Transport across yeast vacuolar and plasma membranes. In The Molecular and Cellular Biology of the Yeast Saccharomyces: Genome Dynamics, Protein Synthesis, and Energetics pp. 523–586 Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  49. Sherman F., Fink G. R., Hicks J. B. 1983 Methods in Yeast Genetics. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  50. Sikorski R. S., Hieter P. 1989; A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. . Genetics 122:19–27
    [Google Scholar]
  51. Singh A., Ursic D., Davies J. 1979; Phenotypic suppression and misreading in Saccharomyces cerevisiae. . Nature 277:146–148
    [Google Scholar]
  52. Strathern J.N.8tHiggins. 1991; Recovery of plasmids from yeast into Escherichia coli: shuttle vectors. Methods Enzymol 194:319–328
    [Google Scholar]
  53. Ulaszewski S., Balzi E., Goffeau A. 1987; Genetic and molecular mapping of the pma1 mutation conferring vanadate resistance to the plasma membrane ATPase from Saccharomyces cerevisiae. . Mol Gen Genet 207:38–46
    [Google Scholar]
  54. Wood K. A., Lord J. M., Wawrzynczak E. J., Piatak M. 1991; Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli. . Eur J Biochem 198:723–732
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-12-3407
Loading
/content/journal/micro/10.1099/13500872-142-12-3407
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error