In vivo identification of Bacillus thuringiensis Cry4Ba toxin receptors by RNA interference knockdown of glycosylphosphatidylinositol-linked aminopeptidase N transcripts in Aedes aegypti larvae.

Autor(es): Saengwiman Suchada; Aroonkesorn Aratee; Dedvisitsakul Plaipol; Sakdee Somsri; Leetachewa Somphob; Angsuthanasombat Chanan; Pootanakit Kusol

Resumo: 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.

Palavras-Chave: Aedes aegypti; Aminopeptidase N; Cry4Ba ? - Endotoxin; RNA interference; Toxin eceptor

Imprenta: Biochemical and Biophysical Research Communications, v. 407, n. 4, p. 708-713, 2011

Identificador do objeto digital: 10.1016/j.bbrc.2011.03.085

Descritores: Aedes aegypti - Molecular Structure ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Proteins ; Aedes aegypti - RNA

Data de publicação: 2011