Cell-cycle-dependent resistance to Bacillus thuringiensis Cry1C toxin in Sf9 cells.
The Sf9 cell line, derived from the moth Spodoptera frugiperda, is highly and specifically sensitive to the Bacillus thuringiensis Cry1C toxin. Upon exposure to Cry1C, ionic pores are formed in the plasma membrane leading to cell swelling and death. Here, we describe a unique transient tolerance to Cry1C of dividing cells, which allowed completion of the division process in the presence of Cry1C. Correlatively, arresting the cells at G2-M phase by nocodazole treatment rendered them insensitive to Cry1C. When the arresting agent was removed, the cells completed their division and gradually regained Cry1C sensitivity. In comparison to normal cells with 1-2% cell-division frequency, the M-phase arrested cells bound less toxin in binding assays. Moreover, no lipid rafts could be isolated from the membranes of M-phase arrested cells. Caveolin-1, identified here for the first time in insect cells, was immunodetected as a lipid raft component of normal cells, but was only present in the membrane-soluble fraction of G2-M-arrested cells. Thus M-phase-linked changes in lipid raft organization may account for diminished Cry1C binding and toxicity. Furthermore, considering the pivotal role of lipid rafts in different cell functions of many cell types, the lack of organized lipid rafts in dividing cells may transiently affect cell susceptibility to pathogens, toxins and other lipid raft-linked functions.
Department of Plant Sciences, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
Dror
D
Avisar
Michal
M
Segal
Baruch
B
Sneh
Aviah
A
Zilberstein
D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000378:metabolism / Q000494:pharmacology*; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004731:Endotoxins / Q000378:metabolism / Q000494:pharmacology*; D016193:G1 Phase / Q000187:drug effects / Q000502:physiology; D016195:G2 Phase / Q000187:drug effects / Q000502:physiology; D006460:Hemolysin Proteins; D007305:Insecticide Resistance; D007306:Insecticides / Q000378:metabolism / Q000494:pharmacology*; D021962:Membrane Microdomains / Q000378:metabolism*; D008938:Mitosis / Q000187:drug effects; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; C030956:insecticidal crystal protein, Bacillus Thuringiensis