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Co-up-regulation of three P450 genes in response to permethrin exposure in permethrin resistant house flies, Musca domestica.

Zhu F, Li T, Zhang L, Liu N - BMC Physiol. (2008)

Bottom Line: The expression of 3 P450 genes, CYP4D4v2, CYP4G2, and CYP6A38, was co-up-regulated by permethrin treatment in permethrin resistant ALHF house flies in a time and dose-dependent manner.Comparison of the deduced protein sequences of these three P450s from resistant ALHF and susceptible aabys and CS house flies revealed identical protein sequences.Genetic linkage analysis located CYP4D4v2 and CYP6A38 on autosome 5, corresponding to the linkage of P450-mediated resistance in ALHF, whereas CYP4G2 was located on autosome 3, where the major insecticide resistance factor(s) for ALHF had been mapped but no P450 genes reported prior to this study.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA. fangzhudy@uky.edu

ABSTRACT

Background: Insects may use various biochemical pathways to enable them to tolerate the lethal action of insecticides. For example, increased cytochrome P450 detoxification is known to play an important role in many insect species. Both constitutively increased expression (overexpression) and induction of P450s are thought to be responsible for increased levels of detoxification of insecticides. However, unlike constitutively overexpressed P450 genes, whose expression association with insecticide resistance has been extensively studied, the induction of P450s is less well characterized in insecticide resistance. The current study focuses on the characterization of individual P450 genes that are induced in response to permethrin treatment in permethrin resistant house flies.

Results: The expression of 3 P450 genes, CYP4D4v2, CYP4G2, and CYP6A38, was co-up-regulated by permethrin treatment in permethrin resistant ALHF house flies in a time and dose-dependent manner. Comparison of the deduced protein sequences of these three P450s from resistant ALHF and susceptible aabys and CS house flies revealed identical protein sequences. Genetic linkage analysis located CYP4D4v2 and CYP6A38 on autosome 5, corresponding to the linkage of P450-mediated resistance in ALHF, whereas CYP4G2 was located on autosome 3, where the major insecticide resistance factor(s) for ALHF had been mapped but no P450 genes reported prior to this study.

Conclusion: Our study provides the first direct evidence that multiple P450 genes are co-up-regulated in permethrin resistant house flies through the induction mechanism, which increases overall expression levels of P450 genes in resistant house flies. Taken together with the significant induction of CYP4D4v2, CYP4G2, and CYP6A38 expression by permethrin only in permethrin resistant house flies and the correlation of the linkage of the genes with resistance and/or P450-mediated resistance in resistant ALHF house flies, this study sheds new light on the functional importance of P450 genes in response to insecticide treatment, detoxification of insecticides, the adaptation of insects to their environment, and the evolution of insecticide resistance.

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Alignment of the nucleotide and deduced amino acid sequences of CYP4G2 in ALHF and aabys house flies. The nucleotide polymorphisms between ALHF and aabys are highlighted and underlined.
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Figure 3: Alignment of the nucleotide and deduced amino acid sequences of CYP4G2 in ALHF and aabys house flies. The nucleotide polymorphisms between ALHF and aabys are highlighted and underlined.

Mentions: The full length of the three putative P450 cDNAs were isolated using 5'and/or 3' RACE with the primers (Table 1) designed based on the 5' and/or 3' end sequences of the putative P450 cDNA fragments. The sequences of the 5' and/or 3' RACE amplified cDNAs overlapped with their corresponding putative P450 cDNA fragments. An entire cDNA fragment for each of the putative P450 genes was subsequently amplified for both ALHF and aabys house flies by PCR using the primer pair (Table 1) synthesized based on the respective 5' and 3' end sequences of each gene. The sequences were named CYP4D4v2 [accession number: EF615001, ], CYP4G2 [accession number: EF615002, ], and CYP6A38 [accession number: EF615003, ] by the P450 nomenclature committee (Dr. D. Nelson, personal communication). The cDNA sequences of CYP4D4v2, CYP4G2, and CYP6A38 have open reading frames of 1515, 1647, and 1500 nucleotides encoding proteins of 505, 549, and 500 residues, respectively. Comparison of the deduced protein sequences of CYP4D4v2, CYP4G2, and CYP6A38 between ALHF and aabys revealed identical protein sequences, although several nucleotide polymorphisms were found in the coding regions of these three genes between the two strains (Figs. 2, 3, 4).


Co-up-regulation of three P450 genes in response to permethrin exposure in permethrin resistant house flies, Musca domestica.

Zhu F, Li T, Zhang L, Liu N - BMC Physiol. (2008)

Alignment of the nucleotide and deduced amino acid sequences of CYP4G2 in ALHF and aabys house flies. The nucleotide polymorphisms between ALHF and aabys are highlighted and underlined.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2567968&req=5

Figure 3: Alignment of the nucleotide and deduced amino acid sequences of CYP4G2 in ALHF and aabys house flies. The nucleotide polymorphisms between ALHF and aabys are highlighted and underlined.
Mentions: The full length of the three putative P450 cDNAs were isolated using 5'and/or 3' RACE with the primers (Table 1) designed based on the 5' and/or 3' end sequences of the putative P450 cDNA fragments. The sequences of the 5' and/or 3' RACE amplified cDNAs overlapped with their corresponding putative P450 cDNA fragments. An entire cDNA fragment for each of the putative P450 genes was subsequently amplified for both ALHF and aabys house flies by PCR using the primer pair (Table 1) synthesized based on the respective 5' and 3' end sequences of each gene. The sequences were named CYP4D4v2 [accession number: EF615001, ], CYP4G2 [accession number: EF615002, ], and CYP6A38 [accession number: EF615003, ] by the P450 nomenclature committee (Dr. D. Nelson, personal communication). The cDNA sequences of CYP4D4v2, CYP4G2, and CYP6A38 have open reading frames of 1515, 1647, and 1500 nucleotides encoding proteins of 505, 549, and 500 residues, respectively. Comparison of the deduced protein sequences of CYP4D4v2, CYP4G2, and CYP6A38 between ALHF and aabys revealed identical protein sequences, although several nucleotide polymorphisms were found in the coding regions of these three genes between the two strains (Figs. 2, 3, 4).

Bottom Line: The expression of 3 P450 genes, CYP4D4v2, CYP4G2, and CYP6A38, was co-up-regulated by permethrin treatment in permethrin resistant ALHF house flies in a time and dose-dependent manner.Comparison of the deduced protein sequences of these three P450s from resistant ALHF and susceptible aabys and CS house flies revealed identical protein sequences.Genetic linkage analysis located CYP4D4v2 and CYP6A38 on autosome 5, corresponding to the linkage of P450-mediated resistance in ALHF, whereas CYP4G2 was located on autosome 3, where the major insecticide resistance factor(s) for ALHF had been mapped but no P450 genes reported prior to this study.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA. fangzhudy@uky.edu

ABSTRACT

Background: Insects may use various biochemical pathways to enable them to tolerate the lethal action of insecticides. For example, increased cytochrome P450 detoxification is known to play an important role in many insect species. Both constitutively increased expression (overexpression) and induction of P450s are thought to be responsible for increased levels of detoxification of insecticides. However, unlike constitutively overexpressed P450 genes, whose expression association with insecticide resistance has been extensively studied, the induction of P450s is less well characterized in insecticide resistance. The current study focuses on the characterization of individual P450 genes that are induced in response to permethrin treatment in permethrin resistant house flies.

Results: The expression of 3 P450 genes, CYP4D4v2, CYP4G2, and CYP6A38, was co-up-regulated by permethrin treatment in permethrin resistant ALHF house flies in a time and dose-dependent manner. Comparison of the deduced protein sequences of these three P450s from resistant ALHF and susceptible aabys and CS house flies revealed identical protein sequences. Genetic linkage analysis located CYP4D4v2 and CYP6A38 on autosome 5, corresponding to the linkage of P450-mediated resistance in ALHF, whereas CYP4G2 was located on autosome 3, where the major insecticide resistance factor(s) for ALHF had been mapped but no P450 genes reported prior to this study.

Conclusion: Our study provides the first direct evidence that multiple P450 genes are co-up-regulated in permethrin resistant house flies through the induction mechanism, which increases overall expression levels of P450 genes in resistant house flies. Taken together with the significant induction of CYP4D4v2, CYP4G2, and CYP6A38 expression by permethrin only in permethrin resistant house flies and the correlation of the linkage of the genes with resistance and/or P450-mediated resistance in resistant ALHF house flies, this study sheds new light on the functional importance of P450 genes in response to insecticide treatment, detoxification of insecticides, the adaptation of insects to their environment, and the evolution of insecticide resistance.

Show MeSH