title: Endogenous expression of a Bt toxin receptor in the Cry1Ac-susceptible insect cell line and its synergistic effect with cadherin on cytotoxicity of activated Cry1Ac.
abstract: Although many insect cell lines derived from various tissues are available, it is unclear whether endogenous receptors of Bacillus thuringiensis (Bt) crystal toxins are expressed in these cell lines. In the present study, we demonstrated that the ovaries-derived Spodoptera litura Sl-HP cell line was susceptible to activated Cry1Ac although larvae of S. litura are not susceptible to the toxin. Assays of the transcriptome revealed that thirteen ATP-binding cassette transporter genes (ABC) were expressed at different levels in this cell line. Of these, the SlABCC3 shared 52-55% amino acid sequence identity with the known Bt toxin receptor ABCC2. RNAi-mediated knockdown targeting SlABCC3 significantly decreased the susceptibility of Sl-HP cells to activated Cry1Ac. Over-expression of the gene strongly increased the susceptibility of Trichoplusia ni Hi5 cells to the toxin. Not only was SlABCC3 comparable to the heterologously expressed Helicoverpa armigera Hacadherin on the receptor-mediated cytotoxicity of activated Cry1Ac to Hi5 cells, but also SlABCC3 and Hacadherin had a strong synergistic effect on cytotoxicity of activated Cry1Ac. These results suggested that Bt toxin receptors-expressing insect cell lines can be used as an alternative model for evaluating cytotoxicity of Bt toxins and studying their mechanisms of action.
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Zuwen
authors: Z
authors: Chen
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Fei
authors: F
authors: He
authors: State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China.
authors: Yutao
authors: Y
authors: Xiao
authors: State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China.
authors: Chenxi
authors: C
authors: Liu
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Jianghuai
authors: J
authors: Li
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Yongbo
authors: Y
authors: Yang
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Hui
authors: H
authors: Ai
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Jianxin
authors: J
authors: Peng
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China.
authors: Huazhu
authors: H
authors: Hong
authors: School of Life Sciences, Central China Normal University, Wuhan 430079, PR China. Electronic address: liukaiyu@mail.ccnu.edu.cn.
authors: Kaiyu
authors: K
authors: Liu
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001413:Bacillus thuringiensis / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D001905:Botulinum Toxins / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism* / Q000633:toxicity; D002460:Cell Line; D004357:Drug Synergism; D004731:Endotoxins / Q000378:metabolism* / Q000633:toxicity; D006460:Hemolysin Proteins / Q000378:metabolism* / Q000633:toxicity; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000187:drug effects / Q000378:metabolism; D034622:RNA Interference; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D018411:Spodoptera / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D018528:ATP-Binding Cassette Transporters; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D015820:Cadherins; C095416:Cry toxin receptors; D004731:Endotoxins; D006460:Hemolysin Proteins; D019476:Insect Proteins; D011956:Receptors, Cell Surface; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D001905:Botulinum Toxins
keywords: ATP-binding cassette transporter; Bacillus thuringiensis; Cadherin; Cytotoxicity; Spodoptera litura cell line