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Two lepidopteran cell lines stably transformed by the ABC transporter gene PDR5 show tolerance to diacetoxyscirpenol

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Summary

The pleiotropic drug resistance 5 gene (pdr5) encodes a multidrug membrane transporter and plays a very important role in the efflux of a broad range of chemicals in yeast cells. To study the possible function of pdr5 in insect cells, two stably pdr5-transformed lepidopteran insect cell lines, Sf21 and CF-203, were developed. Transcripts of pdr5 were detected in these two lines using Northern blotting and RT-PCR analysis. When cells were treated with the protein synthesis inhibitor diacetoxyscirpenol, the transformed Sf21 and CF-203 cell lines showed increased tolerance to this chemical. However, unlike in yeast cells, ecdysone agonist RH5992 could not be excluded by PDR5, probably because of low expression levels or imperfect incorporation of the recombinant protein in these transformed cell lines.

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References

  • Balzi, E.; Wang, M.; Leterme, S.; Van Dyck, L.; Goffeau, A. PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1. J. Biol. Chem. 269:2206–2214; 1994.

    PubMed  CAS  Google Scholar 

  • Dhadialla, T. S.; Carlson, G. R.; Le, D. P. New insecticides with ecdysteroidal and juvenile hormone activity. Annu. Rev. Entomol. 43:545–569; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Golin, J.; Ambukdar, S. V.; Gottesman, M. M.; Habib, A. D.; Sczepanski, J. Studies with novel PDR5p substrates demonstrate a strong size dependence for xenobiontic effux. J. Biol. Chem. 278:5963–5969; 2003.

    Article  PubMed  CAS  Google Scholar 

  • Hu, W. Q.; Feng, Q. L.; Palli, S. R.; Krell, P.; Arif, B. M.; Retnakaran, A. The ABC transporter PDR5p mediates the efflux of nonsteroidal ecdysone agonists in Saccharomyces cerevisiae. Eur. J. Biochem. 268:3416–3422; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Kolaczkowski, M.; van der Rest, M.; Cybularz-Kolaczkowska, A. Anticancer drugs, ionophoric peptides, and steroids as substrates of the yeast multidrug transporter PDR5p. J. Biol. Chem. 271:31543–31548; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Mahe, Y.; Lemoine, Y.; Kuchler, K. The ATP binding cassette transporters PDR5 and Snq2 of Saccharomyces cerevisiae can mediate transport of steroids in vivo J. Biol. Chem. 271:25167–25172; 1996.

    Article  PubMed  CAS  Google Scholar 

  • Muhitch, M. J.; McCormick, S. P.; Alexander, N. J.; Hohn, T. M. Transgenic expression of the TRI101 or PDR5 gene increases resistance of tobacco to the phytotoxic effects of the trichothecene 4,15-diacetoxyscirpenol. Plant Science 157:201–207; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Retnakaran, A.; Hiruma, K.; Palli, S. R.; Riddford, L. M. Molecular analysis of the mode of action of RH-5992, a lepidopteran-specific, non-steroidal ecdysteroid agonist. Insect Biochem. Mol. Biol. 25:109–117; 1995.

    Article  CAS  Google Scholar 

  • Retnakaran, A.; Oberlander, H. Control of chitin synthesis in insects. In: Muzarelli, R. A. A., ed. Chitin enzymology. Ancona, Italy: European Chitin Society; 1993:89–99.

    Google Scholar 

  • Slama, K. Hormonal status of RH-5849 and RH-5992 synthetic ecdysone agonists (eodysoids) examined on several standard bioassays for ecdysteroids. Eur. J. Entomol. 92:317–323; 1995.

    CAS  Google Scholar 

  • Smagghe, G.; Degheele, D. Action of a novel nonsteroidal ecdysteroid mimic, tebufenozide (RH-5992), on insects of different orders. Pesticide Science 42:85–92; 1994.

    Article  CAS  Google Scholar 

  • Sohi, S. S.; Lalouette, W.; macDonld, J. A.; Gringorten, J. L.; Budau, C. B. Establishment of continuous midgut cell lines of spruce budworm (Lepidoptera: Tortricidae). In Vitro Cell. Dev. Biol. 29A(Pt. II):56A; 1993.

    Google Scholar 

  • Sundaram, M.; Palli, S. R.; Krell, P.; Sohi, S. S.; Dhadialla, T. S.; Retnakraran, A. Basis for selective action of a synthetic molting hormone agonist, RH-5992 on lepidopteran insects. Insect Biochem. Mol. Biol. 28:693–704; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Ueno, Y.; Nakajima, M.; Sakai, K.; Ishij, K.; Sato, N.; Shimada, N. Comparative toxicology of trichothec mycotoxins: Inhibition of protein synthesis in animal cells. J. Biochem. 74:285–296; 1973.

    PubMed  CAS  Google Scholar 

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Correspondence to Q. -L. Feng.

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Zhang, D.Y., Krell, P.J. & Feng, Q.L. Two lepidopteran cell lines stably transformed by the ABC transporter gene PDR5 show tolerance to diacetoxyscirpenol. In Vitro Cell.Dev.Biol.-Animal 42, 27–32 (2006). https://doi.org/10.1007/s11626-006-0008-8

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  • DOI: https://doi.org/10.1007/s11626-006-0008-8

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