Aedes aegypti Mos20 cells internalizes cry toxins by endocytosis, and actin has a role in the defense against Cry11Aa toxin.
Bacillus thuringiensis (Bt) Cry toxins are used to control Aedes aegypti, an important vector of dengue fever and yellow fever. Bt Cry toxin forms pores in the gut cells, provoking larvae death by osmotic shock. Little is known, however, about the endocytic and/or degradative cell processes that may counteract the toxin action at low doses. The purpose of this work is to describe the mechanisms of internalization and detoxification of Cry toxins, at low doses, into Mos20 cells from A. aegypti, following endocytotic and cytoskeletal markers or specific chemical inhibitors. Here, we show that both clathrin-dependent and clathrin-independent endocytosis are involved in the internalization into Mos20 cells of Cry11Aa, a toxin specific for Dipteran, and Cry1Ab, a toxin specific for Lepidoptera. Cry11Aa and Cry1Ab are not directed to secretory lysosomes. Instead, Mos20 cells use the Rab5 and Rab11 pathways as a common mechanism, most probably for the expulsion of Cry11Aa and Cry1Ab toxins. In conclusion, we propose that endocytosis is a mechanism induced by Cry toxins independently of specificity, probably as part of a basal immune response. We found, however, that actin is necessary for defense-specific response to Cry11Aa, because actin-silenced Mos20 cells become more sensitive to the toxic action of Cry11A toxin. Cry toxin internalization analysis in insect cell lines may contribute to a better understanding to Cry resistance in mosquitoes.
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico. advcab@ibt.unam.mx.
Adriana
A
Vega-Cabrera
Facultad de Ciencias, Universidad Nacional Autónoma de México, Av. Universidad 3000, Coyoacán, Distrito Federal 04510, Mexico;. angelescancino@gmail.com.
Angeles
A
Cancino-Rodezno
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico. helena@ibt.unam.mx.
Helena
H
Porta
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico. liliana@ibt.unam.mx.
Liliana
L
Pardo-Lopez
D000199:Actins / Q000378:metabolism*; D000330:Aedes; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D004731:Endotoxins / Q000633:toxicity*; D006460:Hemolysin Proteins / Q000633:toxicity*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism
D000199:Actins; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D002966:Clathrin; D004731:Endotoxins; D006460:Hemolysin Proteins; D008565:Membrane Proteins; C106354:flotillins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D007770:L-Lactate Dehydrogenase