Petersen R A, Zangerl A R, Berenbaum M R, Schuler M A
Department of Entomology, University of Illinois at Urbana-Champaign, 320 Morrill Hall, 505 S. Goodwin Ave, Urbana, IL 61801, USA.
Insect Biochem Mol Biol. 2001 Apr 27;31(6-7):679-90. doi: 10.1016/s0965-1748(00)00174-0.
The CYP6B1 and CYP6B3 cytochrome P450 monooxygenases in the midgut of the black swallowtail participate in the metabolism of toxic furanocoumarins present in its host plants. In this study, biochemical analyses indicate that the fat body metabolizes significant amounts of the linear furanocoumarins bergapten and xanthotoxin after larvae feed on xanthotoxin. Northern analyses of the combined CYP6B1/3 transcript expression patterns indicate that transcripts in this P450 subfamily are induced in the midgut and fat body by xanthotoxin. Semi-quantitative RT-PCR analyses of individual CYP6B1/CYP6B3 mRNAs indicate that CYP6B1 transcripts are induced by xanthotoxin in all tissues examined and that CYP6B3 transcripts are induced in the fat body only. These results indicate that the fat body participates in the P450-mediated metabolism of excess furanocoumarins unmetabolized by the midgut. Although transcripts of both genes were detected and CYP6B1 transcripts were induced by xanthotoxin in the integument, furanocoumarin metabolism was not detected. Comparison of these P450 promoters with the promoters of alcohol dehydrogenase genes expressed in the fat bodies of several Drosophila species suggest that the xanthotoxin inducibilities of these P450 genes in fat bodies are regulated by elements other than those modulating expression of Adh genes.
黑凤蝶中肠中的CYP6B1和CYP6B3细胞色素P450单加氧酶参与其寄主植物中有毒呋喃香豆素的代谢。在本研究中,生化分析表明,幼虫取食黄毒素后,脂肪体可代谢大量的线性呋喃香豆素佛手柑内酯和黄毒素。对CYP6B1/3转录本联合表达模式的Northern分析表明,该P450亚家族的转录本在中肠和脂肪体中被黄毒素诱导。对单个CYP6B1/CYP6B3 mRNA进行半定量RT-PCR分析表明,CYP6B1转录本在所有检测组织中均被黄毒素诱导,而CYP6B3转录本仅在脂肪体中被诱导。这些结果表明脂肪体参与了由P450介导的、中肠未代谢的过量呋喃香豆素的代谢。虽然在体壁中检测到了这两个基因的转录本且CYP6B1转录本被黄毒素诱导,但未检测到呋喃香豆素的代谢。将这些P450启动子与几种果蝇脂肪体中表达的乙醇脱氢酶基因的启动子进行比较,结果表明,这些P450基因在脂肪体中的黄毒素诱导性受调控于不同于调节Adh基因表达的元件。