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Brief exposures of human body lice to sublethal amounts of ivermectin over-transcribes detoxification genes involved in tolerance.

ABSTRACT: Transcriptional profiling results, using our non-invasive induction assay {short exposure intervals (2-5 h) to sublethal amounts of insecticides [< lethal concentration 3% (LC(3)) at 24 h] administered by stress-reducing means (contact vs. immersion screen) and with induction assessed in a time frame when tolerance is still present [~lethal concentration 90% (LC(90)) in 2-4 h]}, showed that ivermectin-induced detoxification genes from body lice are identified by quantitative real-time PCR analyses. Of the cytochrome P450 monooxygenase and ATP binding cassette transporter genes induced by ivermectin, CYP6CJ1, CYP9AG1, CYP9AG2 and PhABCC4 were respectively most significantly over-expressed, had high basal expression levels and were most closely related to genes from other organisms that metabolized insecticides, including ivermectin. Injection of double-stranded RNAs (dsRNAs) against either CYP9AG2 or PhABCC4 into non-induced female lice reduced their respective transcript level and resulted in increased sensitivity to ivermectin, indicating that these two genes are involved in the xenobiotic metabolism of ivermectin and in the production of tolerance.

SUBMITTER: Yoon KS

PROVIDER: S-EPMC3208734 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature

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Brief exposures of human body lice to sublethal amounts of ivermectin over-transcribes detoxification genes involved in tolerance.

Yoon K S KS Strycharz J P JP Baek J H JH Sun W W Kim J H JH Kang J S JS Pittendrigh B R BR Lee S H SH Clark J M JM

Insect molecular biology 20110907 6

Transcriptional profiling results, using our non-invasive induction assay {short exposure intervals (2-5 h) to sublethal amounts of insecticides [< lethal concentration 3% (LC(3)) at 24 h] administered by stress-reducing means (contact vs. immersion screen) and with induction assessed in a time frame when tolerance is still present [~lethal concentration 90% (LC(90)) in 2-4 h]}, showed that ivermectin-induced detoxification genes from body lice are identified by quantitative real-time PCR analys ...[more]

PMID: 21895817

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Project description:BackgroundCurrent biomass pretreatment by hydrothermal treatment (including acid hydrolysis, steam explosion, and high-temperature steaming) and ionic liquids generally generate inhibitors to the following fermentation process. Furfural is one of the typical inhibitors generated in hydrothermal treatment of biomass. Furfural could inhibit cell growth rate and decrease biofuel productivity of microbes. Candida tropicalis is a promising microbe for the production of biofuels and value-added chemicals using hemicellulose hydrolysate as carbon source. In this study, C. tropicalis showed a comparable ability of furfural tolerance during fermentation. We investigated the mechanism of C. tropicalis's robust tolerance to furfural and relevant metabolic responses to obtain more information for metabolic engineering of microbes for efficient lignocellulose fermentation.ResultsCandida tropicalis showed comparable intrinsic tolerance to furfural and a fast rate of furfural detoxification. C. tropicalis's half maximal inhibitory concentration for furfural with xylose as the sole carbon source was 3.69 g/L, which was higher than that of most wild-type microbes reported in the literature to our knowledge. Even though furfural prolonged the lag phase of C. tropicalis, the final biomass in the groups treated with 1 g/L furfural was slightly greater than that in the control groups. By real-time PCR analysis, we found that the expression of ADH1 in C. tropicalis (ctADH1) was induced by furfural and repressed by ethanol after furfural depletion. The expression of ctADH1 could be regulated by both furfural and ethanol. After the disruption of gene ctADH1, we found that C. tropicalis's furfural tolerance was weakened. To further confirm the function of ctADH1 and enhance Escherichia coli's furfural tolerance, ctADH1 was overexpressed in E. coli BL21 (DE3). The rate of furfural degradation in E. coli BL21 (DE3) with pET-ADH1 (high-copy plasmid) and pCS-ADH1 (medium-copy plasmid) was increased by 1.59-fold and 1.28-fold, respectively.ConclusionsCandida tropicalis was a robust strain with intrinsic tolerance to inhibitor furfural. The mechanism of furfural detoxification and metabolic responses were identified by multiple analyses. Alcohol dehydrogenase 1 was confirmed to be responsible for furfural detoxification. C. tropicalis showed a complex regulation system during furfural detoxification to minimize adverse effects caused by furfural. Furthermore, the mechanism we uncovered in this work was successfully applied to enhance E. coli's furfural tolerance by heterologous expression of ctADH1. The study provides deeper insights into strain modification for biofuel production by efficient lignocellulose fermentation.

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Brief exposures of human body lice to sublethal amounts of ivermectin over-transcribes detoxification genes involved in tolerance.

Publications

Brief exposures of human body lice to sublethal amounts of ivermectin over-transcribes detoxification genes involved in tolerance.

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