Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2003 Aug 1;373(Pt 3):957–963. doi: 10.1042/BJ20030169

Heterologous expression of four glutathione transferase genes genetically linked to a major insecticide-resistance locus from the malaria vector Anopheles gambiae.

Federica Ortelli 1, Louise C Rossiter 1, John Vontas 1, Hilary Ranson 1, Janet Hemingway 1
PMCID: PMC1223529  PMID: 12718742

Abstract

A cluster of eight genes encoding glutathione transferases (GSTs) are located on division 33B of polytene chromosome arm 3R of the African malaria mosquito, Anopheles gambiae. This region of the genome contains a major 1,1,1-trichloro-2,2-bis-( p -chlorophenyl)ethane (DDT)-resistance locus, rtd1. These GSTs belong to the insect-specific Epsilon class and share between 22.6 and 65.2% identity at the amino acid level. Two distinct allelic variants of the Epsilon GST, GSTe1, differing at 12 out of 224 amino acid residues, are present in laboratory and field populations of A. gambiae. To investigate the possible role of these GSTs in conferring resistance to the insecticide DDT, both GSTe1 alleles, plus three additional members of this gene cluster, were expressed in Escherichia coli and the recombinant proteins biochemically characterized. The five putative glutathione transferases encoded catalytically active subunits with variable biochemical properties. For example, the two allelic variants of GSTE1-1 encoded proteins with over 100-fold variation in peroxidase activity, while the three remaining GSTs had no detectable peroxidase activity. Only GSTE2-2 was able to metabolize DDT. Western blots using antibodies raised against these GSTs indicated that the expression of GSTE2-2 is elevated in a DDT-resistant strain of A. gambiae.

Full Text

The Full Text of this article is available as a PDF (186.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997 Sep 1;25(17):3389–3402. doi: 10.1093/nar/25.17.3389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Board P. G., Baker R. T., Chelvanayagam G., Jermiin L. S. Zeta, a novel class of glutathione transferases in a range of species from plants to humans. Biochem J. 1997 Dec 15;328(Pt 3):929–935. doi: 10.1042/bj3280929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Board P. G., Coggan M., Chelvanayagam G., Easteal S., Jermiin L. S., Schulte G. K., Danley D. E., Hoth L. R., Griffor M. C., Kamath A. V. Identification, characterization, and crystal structure of the Omega class glutathione transferases. J Biol Chem. 2000 Aug 11;275(32):24798–24806. doi: 10.1074/jbc.M001706200. [DOI] [PubMed] [Google Scholar]
  4. Daborn P. J., Yen J. L., Bogwitz M. R., Le Goff G., Feil E., Jeffers S., Tijet N., Perry T., Heckel D., Batterham P. A single p450 allele associated with insecticide resistance in Drosophila. Science. 2002 Sep 27;297(5590):2253–2256. doi: 10.1126/science.1074170. [DOI] [PubMed] [Google Scholar]
  5. Dixon D. P., Cole D. J., Edwards R. Dimerisation of maize glutathione transferases in recombinant bacteria. Plant Mol Biol. 1999 Aug;40(6):997–1008. doi: 10.1023/a:1006257305725. [DOI] [PubMed] [Google Scholar]
  6. Habig W. H., Pabst M. J., Jakoby W. B. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974 Nov 25;249(22):7130–7139. [PubMed] [Google Scholar]
  7. Hayes J. D., Pulford D. J. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol. 1995;30(6):445–600. doi: 10.3109/10409239509083491. [DOI] [PubMed] [Google Scholar]
  8. Holt Robert A., Subramanian G. Mani, Halpern Aaron, Sutton Granger G., Charlab Rosane, Nusskern Deborah R., Wincker Patrick, Clark Andrew G., Ribeiro José M. C., Wides Ron. The genome sequence of the malaria mosquito Anopheles gambiae. Science. 2002 Oct 4;298(5591):129–149. doi: 10.1126/science.1076181. [DOI] [PubMed] [Google Scholar]
  9. Huang H. S., Hu N. T., Yao Y. E., Wu C. Y., Chiang S. W., Sun C. N. Molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the diamondback moth, Plutella xylostella. Insect Biochem Mol Biol. 1998 Sep;28(9):651–658. doi: 10.1016/s0965-1748(98)00049-6. [DOI] [PubMed] [Google Scholar]
  10. Mukabayire O., Besansky N. J. Distribution of T1, Q, Pegasus and mariner transposable elements on the polytene chromosomes of PEST, a standard strain of Anopheles gambiae. Chromosoma. 1996 Jun;104(8):585–595. doi: 10.1007/BF00352298. [DOI] [PubMed] [Google Scholar]
  11. Ranson H., Jensen B., Wang X., Prapanthadara L., Hemingway J., Collins F. H. Genetic mapping of two loci affecting DDT resistance in the malaria vector Anopheles gambiae. Insect Mol Biol. 2000 Oct;9(5):499–507. doi: 10.1046/j.1365-2583.2000.00214.x. [DOI] [PubMed] [Google Scholar]
  12. Ranson H., Prapanthadara L. a., Hemingway J. Cloning and characterization of two glutathione S-transferases from a DDT-resistant strain of Anopheles gambiae. Biochem J. 1997 May 15;324(Pt 1):97–102. doi: 10.1042/bj3240097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ranson H., Rossiter L., Ortelli F., Jensen B., Wang X., Roth C. W., Collins F. H., Hemingway J. Identification of a novel class of insect glutathione S-transferases involved in resistance to DDT in the malaria vector Anopheles gambiae. Biochem J. 2001 Oct 15;359(Pt 2):295–304. doi: 10.1042/0264-6021:3590295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ranson Hilary, Claudianos Charles, Ortelli Federica, Abgrall Christelle, Hemingway Janet, Sharakhova Maria V., Unger Maria F., Collins Frank H., Feyereisen René. Evolution of supergene families associated with insecticide resistance. Science. 2002 Oct 4;298(5591):179–181. doi: 10.1126/science.1076781. [DOI] [PubMed] [Google Scholar]
  15. Sheehan D., Meade G., Foley V. M., Dowd C. A. Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J. 2001 Nov 15;360(Pt 1):1–16. doi: 10.1042/0264-6021:3600001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Simmons T. W., Jamall I. S., Lockshin R. A. Selenium modulates peroxidation in the absence of glutathione peroxidase in Musca domestica. Biochem Biophys Res Commun. 1989 Nov 30;165(1):158–163. doi: 10.1016/0006-291x(89)91048-6. [DOI] [PubMed] [Google Scholar]
  17. Snow R. W., Craig M., Deichmann U., Marsh K. Estimating mortality, morbidity and disability due to malaria among Africa's non-pregnant population. Bull World Health Organ. 1999;77(8):624–640. [PMC free article] [PubMed] [Google Scholar]
  18. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vontas J. G., Small G. J., Hemingway J. Glutathione S-transferases as antioxidant defence agents confer pyrethroid resistance in Nilaparvata lugens. Biochem J. 2001 Jul 1;357(Pt 1):65–72. doi: 10.1042/0264-6021:3570065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vontas John G., Small Graham J., Nikou Dimitra C., Ranson Hilary, Hemingway Janet. Purification, molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens. Biochem J. 2002 Mar 1;362(Pt 2):329–337. doi: 10.1042/0264-6021:3620329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vulule J. M., Beach R. F., Atieli F. K., Roberts J. M., Mount D. L., Mwangi R. W. Reduced susceptibility of Anopheles gambiae to permethrin associated with the use of permethrin-impregnated bednets and curtains in Kenya. Med Vet Entomol. 1994 Jan;8(1):71–75. doi: 10.1111/j.1365-2915.1994.tb00389.x. [DOI] [PubMed] [Google Scholar]
  22. Wang J. Y., McCommas S., Syvanen M. Molecular cloning of a glutathione S-transferase overproduced in an insecticide-resistant strain of the housefly (Musca domestica). Mol Gen Genet. 1991 Jun;227(2):260–266. doi: 10.1007/BF00259679. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES