scholarly journals Co-Expression of a Homologous Cytochrome P450 Reductase Is Required for In Vivo Validation of the Tetranychus urticae CYP392A16-Based Abamectin Resistance in Drosophila

Insects ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 829
Author(s):  
Maria Riga ◽  
Aris Ilias ◽  
John Vontas ◽  
Vassilis Douris
Keyword(s):  
Cytochrome P450 ◽  
Tetranychus Urticae ◽  
Transcriptional Analysis ◽  
Pest Species ◽  
Cytochrome P450 Reductase ◽  
Transcription Activator ◽  
P450 Monooxygenase ◽  
P450 Reductase

Overexpression of the cytochrome P450 monooxygenase CYP392A16 has been previously associated with abamectin resistance using transcriptional analysis in the two-spotted spider mite Tetranychus urticae, an important pest species worldwide; however, this association has not been functionally validated in vivo despite the demonstrated ability of CYP392A16 to metabolize abamectin in vitro. We expressed CYP392A16 in vivo via a Gal4 transcription activator protein/Upstream Activating Sequence (GAL4/UAS) system in Drosophila melanogaster flies, driving expression with detoxification tissue-specific drivers. We demonstrated that CYP392A16 expression confers statistically significant abamectin resistance in toxicity bioassays in Drosophila only when its homologous redox partner, cytochrome P450 reductase (TuCPR), is co-expressed in transgenic flies. Our study shows that the Drosophila model can be further improved, to facilitate the functional analysis of insecticide resistance mechanisms acting alone or in combination.

scholarly journals Novel Substrate Specificity and Temperature-Sensitive Activity of Mycosphaerella graminicola CYP51 Supported by the Native NADPH Cytochrome P450 Reductase

2015 ◽  
Vol 81 (10) ◽  
pp. 3379-3386 ◽  
Author(s):  
Claire L. Price ◽  
Andrew G. S. Warrilow ◽  
Josie E. Parker ◽  
Jonathan G. L. Mullins ◽  
W. David Nes ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Mycosphaerella Graminicola ◽  
Temperature Sensitive ◽  
Cytochrome P450 Reductase ◽  
Zymoseptoria Tritici ◽  
Content Type ◽  
P450 Reductase ◽  
Redox Partner

ABSTRACTMycosphaerella graminicola(Zymoseptoria tritici) is an ascomycete filamentous fungus that causes Septoria leaf blotch in wheat crops. In Europe the most widely used fungicides for this major disease are demethylation inhibitors (DMIs). Their target is the essential sterol 14α-demethylase (CYP51), which requires cytochrome P450 reductase (CPR) as its redox partner for functional activity. TheM. graminicolaCPR (MgCPR) is able to catalyze the sterol 14α-demethylation of eburicol and lanosterol when partnered withCandida albicansCYP51 (CaCYP51) and that of eburicol only withM. graminicolaCYP51 (MgCYP51). The availability of the functionalin vivoredox partner enabled thein vitrocatalytic activity of MgCYP51 to be demonstrated for the first time. MgCYP51 50% inhibitory concentration (IC50) studies with epoxiconazole, tebuconazole, triadimenol, and prothioconazole-desthio confirmed that MgCYP51 bound these azole inhibitors tightly. The characterization of the MgCPR/MgCYP51 redox pairing has produced a functional method to evaluate the effects of agricultural azole fungicides, has demonstrated eburicol specificity in the activity observed, and supports the conclusion that prothioconazole is a profungicide.

Abnormal Microsomal Detoxification Implicated in Fanconi Anemia Group C by Interaction of the FAC Protein With NADPH Cytochrome P450 Reductase

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3050-3056 ◽  
Author(s):  
Frank A.E. Kruyt ◽  
Taizo Hoshino ◽  
Johnson M. Liu ◽  
Pius Joseph ◽  
Anil K. Jaiswal ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Fanconi Anemia ◽  
Mammalian Cells ◽  
Flavin Mononucleotide ◽  
Cytochrome P450 Reductase ◽  
Amino Terminal ◽  
Nadph Cytochrome ◽  
P450 Reductase ◽  
Nadph Cytochrome P450 Reductase

The FAC protein encoded by the Fanconi anemia (FA) complementation group C gene is thought to function in the cytoplasm at a step before DNA repair. Because FA cells are susceptible to mitomycin C, we considered the possibility that FAC might interact with enzymes involved in the bioreductive activation of this drug. Here we report that FAC binds to NADPH cytochrome-P450 reductase (RED), a microsomal membrane protein involved in electron transfer, in both transfected COS-1 and normal murine liver cells. FAC-RED interaction requires the amino-terminal region of FAC and the cytosolic, membrane-proximal domain of the reductase. The latter contains a known binding site for flavin mononucleotide (FMN). Addition of FMN to cytosolic lysates disrupts FAC-reductase complexes, while flavin dinucleotide, which binds to a distinct carboxy-terminal domain, fails to alter FAC-RED complexes at concentrations similar to FMN. FAC is also functionally coupled to this enzyme as its expression in COS-1 cells suppresses the ability of RED to reduce cytochrome c in the presence of NADPH. We propose that FAC plays a fundamental role in vivo by attenuating the activity of RED, thereby regulating a major detoxification pathway in mammalian cells. © 1998 by The American Society of Hematology.

Effect of Neurotransmitters on NADPH-Cytochrome P450 Reductase In Vitro Activity

2007 ◽  
Vol 1 (3) ◽  
pp. 172-175 ◽  
Author(s):  
Guillermo Gervasini ◽  
Carmen Martinez ◽  
Julio Benitez ◽  
Jose A.G. Agundez
Keyword(s):  
Cytochrome P450 ◽  
Vitro Activity ◽  
Cytochrome P450 Reductase ◽  
In Vitro Activity ◽  
Nadph Cytochrome ◽  
P450 Reductase ◽  
Nadph Cytochrome P450 Reductase

Abnormal Microsomal Detoxification Implicated in Fanconi Anemia Group C by Interaction of the FAC Protein With NADPH Cytochrome P450 Reductase

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3050-3056 ◽  
Author(s):  
Frank A.E. Kruyt ◽  
Taizo Hoshino ◽  
Johnson M. Liu ◽  
Pius Joseph ◽  
Anil K. Jaiswal ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Fanconi Anemia ◽  
Mammalian Cells ◽  
Flavin Mononucleotide ◽  
Cytochrome P450 Reductase ◽  
Amino Terminal ◽  
Nadph Cytochrome ◽  
P450 Reductase ◽  
Nadph Cytochrome P450 Reductase

Abstract The FAC protein encoded by the Fanconi anemia (FA) complementation group C gene is thought to function in the cytoplasm at a step before DNA repair. Because FA cells are susceptible to mitomycin C, we considered the possibility that FAC might interact with enzymes involved in the bioreductive activation of this drug. Here we report that FAC binds to NADPH cytochrome-P450 reductase (RED), a microsomal membrane protein involved in electron transfer, in both transfected COS-1 and normal murine liver cells. FAC-RED interaction requires the amino-terminal region of FAC and the cytosolic, membrane-proximal domain of the reductase. The latter contains a known binding site for flavin mononucleotide (FMN). Addition of FMN to cytosolic lysates disrupts FAC-reductase complexes, while flavin dinucleotide, which binds to a distinct carboxy-terminal domain, fails to alter FAC-RED complexes at concentrations similar to FMN. FAC is also functionally coupled to this enzyme as its expression in COS-1 cells suppresses the ability of RED to reduce cytochrome c in the presence of NADPH. We propose that FAC plays a fundamental role in vivo by attenuating the activity of RED, thereby regulating a major detoxification pathway in mammalian cells. © 1998 by The American Society of Hematology.

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