Isolation of insecticide resistance-related forms of cytochrome <i>P</i>-450 from <i>Drosophila melanogaster</i>

Sundseth, Scott S.; Nix, Carroll E.; Waters, Larry C.

doi:10.1042/bj2650213

Cited by 29 publications

(5 citation statements)

References 26 publications

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“…The P-450 from phenobarbital-treated rat brain was eluted with low-ionicstrength buffer (10 mm potassium phosphate) from a DEAE-Sephacel column. Similar behaviour has been reported for rabbit lung P-450 [5] and the P-450 from Drosophila melanogaster [33]. SDS/PAGE of purified rat brain P-450 revealed the apparently homogeneous nature of the preparation (Fig.…”

Section: Discussionsupporting

confidence: 84%

Characterization of a phenobarbital-inducible cytochrome P-450, NADPH-cytochrome P-450 reductase and reconstituted cytochrome P-450 mono-oxygenase system from rat brain. Evidence for constitutive presence in rat and human brain

Anandatheerthavarada

Boyd

Ravindranath

1992

Biochemical Journal

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Cytochrome P-450 was purified to apparent homogeneity from the brain microsomes of phenobarbital-treated rats. The specific content of the purified P-450 was 12.7 nmol/mg of protein. NADPH-cytochrome P-450 reductase (reductase) was also purified to apparent homogeneity from brain microsomes. The specific content was 34.7 mumol of cytochrome c reduced/min per mg of protein. The reduced carbon monoxide spectrum of purified P-450 exhibited a peak at 450 nm. Both the P-450 and the reductase moved as single bands on SDS/PAGE. The molecular masses of the purified P-450 and the reductase were determined to be 53.3 and 72.0 kDa respectively. The purified brain P-450 cross-reacted with antibodies to rat liver P-450IIB1/IIB2 when examined by Western immunoblotting, but no immunological similarity was observed with rat liver P-450IA1/IA2 or P-450IIE1. Purified rat brain reductase cross-reacted with antibodies to rat liver reductase. Further, immunoblot experiments with untreated rat and human brain microsomes using antisera to the purified rat brain P-450 and reductase indicated that these forms of P-450 and NADPH-cytochrome P-450 reductase exist constitutively in rat and human brain. Purified rat brain P-450 was reconstituted with purified NADPH-cytochrome P-450 reductase, deoxycholate and dilauroyl glyceryl 3-phosphocholine. NADPH-dependent N-demethylation of aminopyrine and morphine was observed in the reconstituted system. The catalytic-centre activities were 80.25 and 38.2 nmol of formaldehyde formed/min per nmol of P-450 respectively. The reconstituted system had a comparatively lower catalytic-centre activity for 7-ethoxycoumarin O-de-ethylase (10.5 nmol of product formed/min per nmol of P-450).

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Section: Discussionsupporting

confidence: 84%

Characterization of a phenobarbital-inducible cytochrome P-450, NADPH-cytochrome P-450 reductase and reconstituted cytochrome P-450 mono-oxygenase system from rat brain. Evidence for constitutive presence in rat and human brain

Anandatheerthavarada

Boyd

Ravindranath

1992

Biochemical Journal

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“…Resistance to DDT could result from mechanisms other than target site insensitivity. For example, increased mixed function oxidase activity is also known to confer resistance to a wide range of insecticides in some strains of Drosophila (Waters and Nix 1988;Sundseth et al 1990) and DDT dehydrochlorinase may also be responsible for DDT metabolism. Several observations indicate that it is the para mutation that confers DDT resistance in our strains, rather than genetic polymorphisms in the background associated with increased metabolic activity.…”

Section: Ddt and Pyrethroid Resistance Of¯ies Bearing Para Ts Allelesmentioning

confidence: 99%

Point mutations in the Drosophila sodium channel gene para associated with resistance to DDT and pyrethroid insecticides

et al. 1997

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The gene para in Drosophila melanogaster encodes an alpha subunit of voltage-activated sodium channels, the presumed site of action of DDT and pyrethroid insecticides. We used an existing collection of Drosophila para mutants to examine the molecular basis of targetsite resistance to pyrethroids and DDT. Six out of thirteen mutants tested were associated with a largely dominant, 10- to 30-fold increase in DDT resistance. The amino acid lesions associated with these alleles defined four sites in the sodium channel polypeptide where a mutational change can cause resistance: within the intracellular loop between S4 and S5 in homology domains I and III, within the pore region of homology domain III, and within S6 in homology domain III. Some of these sites are analogous with those defined by knockdown resistance (kdr) and super-kdr resistance-associated mutations in houseflies and other insects, but are located in different homologous units of the channel polypeptide. We find a striking synergism in resistance levels with particular heterozygous combinations of para alleles that appears to mimic the super-kdr double mutant housefly phenotype. Our results indicate that the alleles analyzed from natural populations represent only a subset of mutations that can confer resistance. The implications for the binding site of pyrethroids and mechanisms of target-site insensitivity are discussed.

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“…Exposure to a substrate or other inducer may result in elevated levels of a particular isozyme or elevated activity or both (20)(21)(22)(23). In Drosophila, only the cytochrome P450 system of D. melanogaster has been studied, and the emphasis for that species has been on pheromone (ecdysone) processing (24,25), insecticide resistance, and mutagen metabolism and activation (14)(15)(16)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38) instant Drosophila medium at ambient temperature and humidity. Larval induction was accomplished by adding 1.0 g of powdered cactus tissue to the medium surface, rehydrated by adding deionized water.…”

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confidence: 99%

Involvement of cytochrome P450 in host-plant utilization by Sonoran Desert Drosophila.

Frank

Fogleman

1992

Proc. Natl. Acad. Sci. U.S.A.

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The four Drosophila species endemic to the Sonoran Desert (Drosophila mettleri, Drosophila mojavensis, The four species are not closely related phylogenetically, although all are members ofthe major virilis-repleta radiation of the genus Drosophila. These species appear to have adapted independently to the desert environment (2). The major population center of the nearest relative of each species is found outside the desert or overlaps only slightly with the desert. Each of the endemic species also uses a different cactus species as a host, in a nearly one-to-one relationship, as demonstrated by extensive rearing records and collection of adults from naturally occurring rots (1). In any particular location, each Drosophila species generally utilizes the necrotic tissue of only one cactus species. Due to differences in the geographic distribution of the cacti and one case of behavioral preference, three of the four fly species exhibit a shift in host plants between the Baja Peninsula and the mainland Sonoran Desert. D. nigrospiracula uses saguaro (Carnegiea gigantea) on the mainland and card6n (Pachycereus pringlei) on the peninsula. Saguaro cactus is only found on the mainland, and card6n is primarily restricted to the Baja peninsula. The two cacti are morphologically and chemically very similar and have low concentrations (1% dry weight) of simple isoquinoline alkaloids (e.g., carnegine and gigantine in saguaro) as their primary allelochemicals. The host plants for D. mojavensis are agria (Stenocereus gummosus) and organ pipe (Stenocereus thurberi), which are sympatric on the peninsula. D. mojavensis shows a clear preference for agria and utilizes organ pipe on the mainland, where agria is all but absent. Agria and organ pipe do not contain alkaloids, but both have sterol diols, medium-chain

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Isolation of insecticide resistance-related forms of cytochrome P-450 from Drosophila melanogaster

Cited by 29 publications

References 26 publications

Characterization of a phenobarbital-inducible cytochrome P-450, NADPH-cytochrome P-450 reductase and reconstituted cytochrome P-450 mono-oxygenase system from rat brain. Evidence for constitutive presence in rat and human brain

Characterization of a phenobarbital-inducible cytochrome P-450, NADPH-cytochrome P-450 reductase and reconstituted cytochrome P-450 mono-oxygenase system from rat brain. Evidence for constitutive presence in rat and human brain

Point mutations in the Drosophila sodium channel gene para associated with resistance to DDT and pyrethroid insecticides

Involvement of cytochrome P450 in host-plant utilization by Sonoran Desert Drosophila.

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