title: In vivo identification of Bacillus thuringiensis Cry4Ba toxin receptors by RNA interference knockdown of glycosylphosphatidylinositol-linked aminopeptidase N transcripts in Aedes aegypti larvae.
abstract: Bacillus thuringiensis Cry4Ba toxin selectively kills Aedes aegypti mosquito larvae as it is in part due to the presence of specific membrane-bound protein receptors. In this study, using data mining approach, we initially identified three potential glycosylphosphatidylinositol-linked aminopeptidase N (GPI-APN) isoforms, APN2778, APN2783 and APN5808, which are believed to act as Cry4Ba toxin receptors. These three isoforms that are functionally expressed in the larval midgut can be sequence-specific knocked down (ranging from ∼80 % to 95 %) by soaking the Aedes aegypti larvae in buffer of long double-stranded GPI-APN RNAs (∼300-680 bp). Finally, to see the physiological effect of APN knockdowns, the larvae were fed with Escherichia coli expressing Cry4Ba toxin. The results revealed that all the three identified GPI-APN isoforms may possibly function as a Cry4Ba receptor, particularly for APN2783 as those larvae with this transcript knockdown showed a dramatic increase in resistance to Cry4Ba toxicity.
authors: Bacterial Protein Toxin Research Cluster, Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Nakornpathom 73170, Thailand.
authors: Suchada
authors: S
authors: Saengwiman
authors: Aratee
authors: A
authors: Aroonkesorn
authors: Plaipol
authors: P
authors: Dedvisitsakul
authors: Somsri
authors: S
authors: Sakdee
authors: Somphob
authors: S
authors: Leetachewa
authors: Chanan
authors: C
authors: Angsuthanasombat
authors: Kusol
authors: K
authors: Pootanakit
mesh_terms: D000330:Aedes / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000633:toxicity*; D018826:CD13 Antigens / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics*; D004731:Endotoxins / Q000633:toxicity*; D055785:Gene Knockdown Techniques; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006460:Hemolysin Proteins / Q000633:toxicity*; D007306:Insecticides / Q000633:toxicity*; D007527:Isoenzymes / Q000235:genetics; D007814:Larva / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D034622:RNA Interference; D012333:RNA, Messenger / Q000037:antagonists & inhibitors; D020539:Sequence Analysis, Protein; D014158:Transcription, Genetic
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D017261:Glycosylphosphatidylinositols; D006460:Hemolysin Proteins; D007306:Insecticides; D007527:Isoenzymes; D012333:RNA, Messenger; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D018826:CD13 Antigens