High expression of Cyp6g1, a cytochrome P450 gene, does not necessarily confer DDT resistance in Drosophila melanogaster

Srilalitha Kuruganti; Vita Lam; Xuguo Zhou; Gary Bennett; Barry R. Pittendrigh; Ranjan Ganguly

doi:10.1016/j.gene.2006.09.019

High expression of Cyp6g1, a cytochrome P450 gene, does not necessarily confer DDT resistance in Drosophila melanogaster

Srilalitha Kuruganti, Vita Lam, Xuguo Zhou, Gary Bennett, Barry R. Pittendrigh, Ranjan Ganguly

Entomology

Research output: Contribution to journal › Article › peer-review

31 Scopus citations

Abstract

Cytochrome P450 monooxygenases, a family of detoxifying enzymes, are thought to confer resistance to various insecticides including DDT. Daborn et al. [Daborn, P., Yen, J.L., Bogwitz, M., Le Goff, G., Feil, et al. 2002. A single p450 allele associated with insecticide resistance in Drosophila. Science 297, 2253-2256.] suggested that the Accord transposable element causes overexpression of a Cyp6g1 allele, which has spread globally and is the basis of DDT resistance in Drosophila melanogaster populations. To determine whether the same phenomenon also operates in other Drosophila strains, we investigated 91-R, 91-C, ry⁵⁰⁶, Wisconsin, Canton-SH and Hikone-RH strains. While the LC₅₀ values for the 91-R and Wisconsin strains are 8348 μg and 447 μg of DDT, respectively, values for the other four strains range between 0.74 to 20.9 μg. As expected, the susceptible ry⁵⁰⁶ and 91-C strains have about 16-33-fold lower levels of CYP6G1 mRNA than the resistant 91-R and Wisconsin strains. Surprisingly, CYP6G1 mRNA and protein levels in the Canton-SH and Hikone-RH strains are as high as in the two resistant strains, yet they are as susceptible as the 91-C strain. The susceptible phenotype of the Canton-SH and Hikone-RH strains is not due to mutation in the Cyp6g1 gene; sequence analysis showed that Cyp6g1 alleles of resistant and susceptible strains are very similar and cannot be classified into resistant and susceptible alleles. As observed by others, we also found that only the 5′-upstream DNA of overexpressing alleles of Cyp6g1 has an insertional DNA, which is similar to Accord and Ninja elements. To examine the role of Cyp6g1 in DDT resistance, we substituted the Cyp6g1 allele of the 91-R strain with the allele from the susceptible 91-C strain via recombination and synthesized three recombinant lines. All three lines lacked Accord insertion and showed low expression of Cyp6g1 like the 91-C strain, yet they were as highly resistant as the 91-R strain. We conclude a strain may not have to have Accord insertion in the Cyp6g1 gene and the Cyp6g1 itself may not have to be overexpressed for DDT resistance to occur.

Original language	English
Pages (from-to)	43-53
Number of pages	11
Journal	Gene
Volume	388
Issue number	1-2
DOIs	https://doi.org/10.1016/j.gene.2006.09.019
State	Published - Feb 15 2007

Bibliographical note

Funding Information:
The loading control antibody, JLA 20 (actin), was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242. The authors would like to thank Charity Olson for technical assistance and Dr. Arnold Saxton for his advice on probit analysis in SAS. This work was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2002-35302-12281 grant to RG and a National Institutes of Health (NIH 1R01 AI51513-01) grant to BRP. This is publication No. 2005-17477, Purdue University Agricultural Experimental Station, West Lafayette, IN, U.S.A. A major portion of this work will be submitted by SK in partial fulfillment of the requirements for the degree of Doctor of Philosophy.

Keywords

Accord/Ninja insertion
Allele substitution
DDT-resistance assay
DNA sequencing
Northern blot analysis
Western blotting

ASJC Scopus subject areas

Genetics

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10.1016/j.gene.2006.09.019

Cite this

@article{7085d3bc8eef47ba94943f57361f81b8,

title = "High expression of Cyp6g1, a cytochrome P450 gene, does not necessarily confer DDT resistance in Drosophila melanogaster",

abstract = "Cytochrome P450 monooxygenases, a family of detoxifying enzymes, are thought to confer resistance to various insecticides including DDT. Daborn et al. [Daborn, P., Yen, J.L., Bogwitz, M., Le Goff, G., Feil, et al. 2002. A single p450 allele associated with insecticide resistance in Drosophila. Science 297, 2253-2256.] suggested that the Accord transposable element causes overexpression of a Cyp6g1 allele, which has spread globally and is the basis of DDT resistance in Drosophila melanogaster populations. To determine whether the same phenomenon also operates in other Drosophila strains, we investigated 91-R, 91-C, ry506, Wisconsin, Canton-SH and Hikone-RH strains. While the LC50 values for the 91-R and Wisconsin strains are 8348 μg and 447 μg of DDT, respectively, values for the other four strains range between 0.74 to 20.9 μg. As expected, the susceptible ry506 and 91-C strains have about 16-33-fold lower levels of CYP6G1 mRNA than the resistant 91-R and Wisconsin strains. Surprisingly, CYP6G1 mRNA and protein levels in the Canton-SH and Hikone-RH strains are as high as in the two resistant strains, yet they are as susceptible as the 91-C strain. The susceptible phenotype of the Canton-SH and Hikone-RH strains is not due to mutation in the Cyp6g1 gene; sequence analysis showed that Cyp6g1 alleles of resistant and susceptible strains are very similar and cannot be classified into resistant and susceptible alleles. As observed by others, we also found that only the 5′-upstream DNA of overexpressing alleles of Cyp6g1 has an insertional DNA, which is similar to Accord and Ninja elements. To examine the role of Cyp6g1 in DDT resistance, we substituted the Cyp6g1 allele of the 91-R strain with the allele from the susceptible 91-C strain via recombination and synthesized three recombinant lines. All three lines lacked Accord insertion and showed low expression of Cyp6g1 like the 91-C strain, yet they were as highly resistant as the 91-R strain. We conclude a strain may not have to have Accord insertion in the Cyp6g1 gene and the Cyp6g1 itself may not have to be overexpressed for DDT resistance to occur.",

keywords = "Accord/Ninja insertion, Allele substitution, DDT-resistance assay, DNA sequencing, Northern blot analysis, Western blotting",

author = "Srilalitha Kuruganti and Vita Lam and Xuguo Zhou and Gary Bennett and Pittendrigh, {Barry R.} and Ranjan Ganguly",

note = "Funding Information: The loading control antibody, JLA 20 (actin), was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242. The authors would like to thank Charity Olson for technical assistance and Dr. Arnold Saxton for his advice on probit analysis in SAS. This work was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2002-35302-12281 grant to RG and a National Institutes of Health (NIH 1R01 AI51513-01) grant to BRP. This is publication No. 2005-17477, Purdue University Agricultural Experimental Station, West Lafayette, IN, U.S.A. A major portion of this work will be submitted by SK in partial fulfillment of the requirements for the degree of Doctor of Philosophy.",

year = "2007",

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doi = "10.1016/j.gene.2006.09.019",

language = "English",

volume = "388",

pages = "43--53",

number = "1-2",

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TY - JOUR

T1 - High expression of Cyp6g1, a cytochrome P450 gene, does not necessarily confer DDT resistance in Drosophila melanogaster

AU - Kuruganti, Srilalitha

AU - Lam, Vita

AU - Zhou, Xuguo

AU - Bennett, Gary

AU - Pittendrigh, Barry R.

AU - Ganguly, Ranjan

N1 - Funding Information: The loading control antibody, JLA 20 (actin), was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242. The authors would like to thank Charity Olson for technical assistance and Dr. Arnold Saxton for his advice on probit analysis in SAS. This work was supported by the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2002-35302-12281 grant to RG and a National Institutes of Health (NIH 1R01 AI51513-01) grant to BRP. This is publication No. 2005-17477, Purdue University Agricultural Experimental Station, West Lafayette, IN, U.S.A. A major portion of this work will be submitted by SK in partial fulfillment of the requirements for the degree of Doctor of Philosophy.

PY - 2007/2/15

Y1 - 2007/2/15

N2 - Cytochrome P450 monooxygenases, a family of detoxifying enzymes, are thought to confer resistance to various insecticides including DDT. Daborn et al. [Daborn, P., Yen, J.L., Bogwitz, M., Le Goff, G., Feil, et al. 2002. A single p450 allele associated with insecticide resistance in Drosophila. Science 297, 2253-2256.] suggested that the Accord transposable element causes overexpression of a Cyp6g1 allele, which has spread globally and is the basis of DDT resistance in Drosophila melanogaster populations. To determine whether the same phenomenon also operates in other Drosophila strains, we investigated 91-R, 91-C, ry506, Wisconsin, Canton-SH and Hikone-RH strains. While the LC50 values for the 91-R and Wisconsin strains are 8348 μg and 447 μg of DDT, respectively, values for the other four strains range between 0.74 to 20.9 μg. As expected, the susceptible ry506 and 91-C strains have about 16-33-fold lower levels of CYP6G1 mRNA than the resistant 91-R and Wisconsin strains. Surprisingly, CYP6G1 mRNA and protein levels in the Canton-SH and Hikone-RH strains are as high as in the two resistant strains, yet they are as susceptible as the 91-C strain. The susceptible phenotype of the Canton-SH and Hikone-RH strains is not due to mutation in the Cyp6g1 gene; sequence analysis showed that Cyp6g1 alleles of resistant and susceptible strains are very similar and cannot be classified into resistant and susceptible alleles. As observed by others, we also found that only the 5′-upstream DNA of overexpressing alleles of Cyp6g1 has an insertional DNA, which is similar to Accord and Ninja elements. To examine the role of Cyp6g1 in DDT resistance, we substituted the Cyp6g1 allele of the 91-R strain with the allele from the susceptible 91-C strain via recombination and synthesized three recombinant lines. All three lines lacked Accord insertion and showed low expression of Cyp6g1 like the 91-C strain, yet they were as highly resistant as the 91-R strain. We conclude a strain may not have to have Accord insertion in the Cyp6g1 gene and the Cyp6g1 itself may not have to be overexpressed for DDT resistance to occur.

AB - Cytochrome P450 monooxygenases, a family of detoxifying enzymes, are thought to confer resistance to various insecticides including DDT. Daborn et al. [Daborn, P., Yen, J.L., Bogwitz, M., Le Goff, G., Feil, et al. 2002. A single p450 allele associated with insecticide resistance in Drosophila. Science 297, 2253-2256.] suggested that the Accord transposable element causes overexpression of a Cyp6g1 allele, which has spread globally and is the basis of DDT resistance in Drosophila melanogaster populations. To determine whether the same phenomenon also operates in other Drosophila strains, we investigated 91-R, 91-C, ry506, Wisconsin, Canton-SH and Hikone-RH strains. While the LC50 values for the 91-R and Wisconsin strains are 8348 μg and 447 μg of DDT, respectively, values for the other four strains range between 0.74 to 20.9 μg. As expected, the susceptible ry506 and 91-C strains have about 16-33-fold lower levels of CYP6G1 mRNA than the resistant 91-R and Wisconsin strains. Surprisingly, CYP6G1 mRNA and protein levels in the Canton-SH and Hikone-RH strains are as high as in the two resistant strains, yet they are as susceptible as the 91-C strain. The susceptible phenotype of the Canton-SH and Hikone-RH strains is not due to mutation in the Cyp6g1 gene; sequence analysis showed that Cyp6g1 alleles of resistant and susceptible strains are very similar and cannot be classified into resistant and susceptible alleles. As observed by others, we also found that only the 5′-upstream DNA of overexpressing alleles of Cyp6g1 has an insertional DNA, which is similar to Accord and Ninja elements. To examine the role of Cyp6g1 in DDT resistance, we substituted the Cyp6g1 allele of the 91-R strain with the allele from the susceptible 91-C strain via recombination and synthesized three recombinant lines. All three lines lacked Accord insertion and showed low expression of Cyp6g1 like the 91-C strain, yet they were as highly resistant as the 91-R strain. We conclude a strain may not have to have Accord insertion in the Cyp6g1 gene and the Cyp6g1 itself may not have to be overexpressed for DDT resistance to occur.

KW - Accord/Ninja insertion

KW - Allele substitution

KW - DDT-resistance assay

KW - DNA sequencing

KW - Northern blot analysis

KW - Western blotting

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High expression of Cyp6g1, a cytochrome P450 gene, does not necessarily confer DDT resistance in Drosophila melanogaster

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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