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Genetic and molecular evidence for a trans-acting regulatory locus controlling glutathione S-transferase-2 expression in Aedes aegypti

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Summary

The amount of glutathione S-transferase-2 (GST-2) protein and enzyme activity in a mutant strain (strain GG) of the yellow fever mosquito (Aedes aegypti) is approximately 25-fold higher than in the wild-type (+ +) strain. The mode of inheritance of the GG phenotype was studied in F1 and backcross progeny using GST enzyme assays, isozyme-specific antisera, and Northern blot analysis. Enzyme assay of parental and F1 progeny showed that the + + phenotype was dominant to the GG phenotype. This was true for larvae as well as for all tissues examined in adults in both sexes. Immunoblotting experiments showed that, like the + + strain, F1 larvae and adults express very low levels of GST-2 protein compared with the GG strain. Northern blotting experiments showed that the steady-state levels of GST-2 mRNA in parental and F1 hybrid larvae closely matched the enzyme activity and immunological data. These results suggest the existence of a trans-acting regulatory locus that acts to repress GST-2 mRNA transcription and/or decrease GST-2 mRNA stability in + + and F1 hybrids. GST enzyme activity in backcross progeny, however, did not segregate into the two distinct phenotypes (low and high) predicted for a single locus, dominant allele model. Backcross progeny expressed a wide range of GST activity and GST-2 protein amount with no apparent fit to simple Mendelian ratios. These backcross data suggest that additional loci are also involved in regulating GST-2 isozyme expression. Taken together, the results suggest that overexpression of GST-2 in the GG strain of Aedes aegypti is due to allelic segregation at a trans-acting regulatory locus, most likely a repressor, with additional effects due to other modifying loci.

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References

  • Batist G, Tulpule A, Sinha BK, Katki AG, Myers CE, Cowan KH (1986) Overexpression of a novel anionic glutathione transferase in multidrug-resistant human breast cancer cells. J Biol Chem 261:15544–15549

    Google Scholar 

  • Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC (1984) A rapid, sensitive method for detection of alkaline phosphataseconjugated anti-antibody on western blots. Anal Biochem 136:175–179

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203

    Google Scholar 

  • Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159

    Article  CAS  PubMed  Google Scholar 

  • Clark AG, Shamaan NA (1984) Evidence that DDT-dehydrochlorinase from the house fly is a glutathione S-transferase. Pest Biochem Physiol 22:249–261

    Google Scholar 

  • Corbin V, Maniatis T (1990) Identification of cis-regulatory elements required for larval expression of the Drosophila melanogaster alcohol dehydrogenase gene. Genetics 124:637–646

    Google Scholar 

  • Devonshire AL, Moores GD (1982) A carboxylesterase with broad substrate specificity causes organophosphorus, carbamate, and pyrethroid resistance in peach-potato aphids (Myzus persicae). Pest Biochem Physiol 18:235–246

    Google Scholar 

  • Dickinson WJ (1980) Tissue specificity of enzyme expression regulated by diffusible factors: Evidence in Drosophila hybrids. Science 207:29

    Google Scholar 

  • Dickinson WJ (1983) Tissue-specific allelic isozyme patterns and cis-acting development regulators. In: Rattazzi MC, Scandalios JG, Whitt GS (eds) Isozymes. Current topics in biological and medical research, vol 9: Gene expression and development. AR hiss, New York, pp 107–122

    Google Scholar 

  • Grant DF (1991) Evolution of glutathione S-transferase subunits in Culicidae and related Nematocera: Electrophoretic and immunological evidence for conserved enzyme structure and expression. Insect Biochem 21:435–445

    Google Scholar 

  • Grant DF, Matsumura F (1988) Glutathione S-transferase-1 in Aedes aegypti larvae: purification and properties. Insect Biochem 18:615–622

    Google Scholar 

  • Grant DF, Matsumura F (1989) Glutathione S-transferase 1 and 2 in susceptible and insecticide-resistant Aedes aegypti. Pest Biochem Physiol 33:132–143

    Google Scholar 

  • Grant DF, Bender DM, Hammock BD (1989) Quantitative kinetic assays for glutathione S-transferase and general esterase in individual mosquitoes using an EIA reader. Insect Biochem 19:741–751

    Google Scholar 

  • Grant GF, Dietze EC, Hammock BD (1991) Glutathione S-transferase isozymes in Aedes aegypti: purfication, characterization, and isozyme-specific regulation. Insect Biochem 21:421–433

    Google Scholar 

  • Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases, the first enzymatic step in mercapturin acid formation. J Biol Chem 249:7130–7139

    Google Scholar 

  • Hall BG (1983) Evolution of new metabolic functions in laboratory organisms. In: Evolution of genes and proteins. Nei M, Kohen RK (eds) Sinauer Sunderland, England, pp 234–257

    Google Scholar 

  • Hall BG, Hard DL (1975) Regulation of newly evolved enzymes. II. The ebg repressor. Genetics 81:427–435

    Google Scholar 

  • Hemingway J, Miyamoto J, Herath PRJ (1991) A possible novel link between organophosphorus and DDT insecticide resistance genes in Anopheles: Supporting evidence from fenitrothion metabolism studies. Pest Biochem Physiol 39:49–56

    Google Scholar 

  • Jansen R, Ledley FD (1989) Production of discrete high-specific activity DNA probes using the polymerase chain reaction. Gene Anal Techniques 6:79–83

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    PubMed  Google Scholar 

  • Lem J, Chin AC, Thayer MJ, Leach RJ, Fournier REK (1988) Coordinate regulation of two genes encoding gluconeogenic enzymes by the trans-dominant locus Tse-1. Proc Natl Acad Sci USA 85:7302–7306

    Google Scholar 

  • Mankowitz L, Castro VM, Mannervik B, Rydstrom J, DePierre JW (1990) Increase in the amount of glutathione transferase 4-4 in the rat adrenal gland after hypophysectomy and down-regulation by subsequent treatment with adrenocorticotrophic hormone. Biochem J 265:147–154

    Google Scholar 

  • Mouches C, Pasteru N, Berge JB, Hyrien O, Raymond M, De Saint Vincent BR, De Silvestri M, Georghiou GP (1986) Amplification of an esterase gene is responsible for insecticide resistance in a California Culex mosquito. Science 233:778–780

    Google Scholar 

  • Munstermann LE, Craig GB (1979) Genetics of Aedes aegypti: updating the linkage map. J Hered 70:291–296

    Google Scholar 

  • Pickett CB, Lu AYH (1989) Glutathione S-transferases: gene structure, regulation, and biological function. Annu Rev Biochem 58:743–764

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Scott JG, Lee SST, Shono T (1990) Biochemical changes in the cytochrome P450 monooxygenases of seven insecticide-resistant house fly (Musca domestica L.) strains. Pest Biochem Physiol 36:127–134

    Google Scholar 

  • Sokal RR, Rohlf FJ (1969) Biometry. W.H. Freeman and Company, San Francisco, 776

    Google Scholar 

  • Telakowski-Hopkins CA, King RG, Pickett CB (1988) Glutathione S-transferase Ya subunit gene: identification of regulatory elements required for basal level and inducible expression. Proc Natl Acad Sci USA 85:1000–1004

    Google Scholar 

  • Wang J-Y, McCommas S, Syvanen M (1991) Molecular cloning of a glutathione S-transferase overproduced in an insecticide-resistant strain of the housefly (Musca domestica). Mol Gen Genet 227:260–266

    Google Scholar 

  • Wu C-Y, Mote J Jr, Brennan MD (1990) Tissue-specific expression phenotypes of Hawaiian Drosophila Adh genes in Drosophila melanogaster transformants. Genetics 125:599–610

    Google Scholar 

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Communicated by D. Finnegan

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Grant, D.F., Hammock, B.D. Genetic and molecular evidence for a trans-acting regulatory locus controlling glutathione S-transferase-2 expression in Aedes aegypti . Molec. Gen. Genet. 234, 169–176 (1992). https://doi.org/10.1007/BF00283836

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  • DOI: https://doi.org/10.1007/BF00283836

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