CulturedAedes albopictus mosquito cells synthesize hormone-inducible proteins |
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Authors: | Que Lan Anna Gerenday Ann Marie Fallon |
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Institution: | (1) Department of Entomology, University of Minnesota, 1980 Folwell Ave., 55108 St. Paul, Minnesota;(2) Present address: Department of Zoology, University of Washington, 98195 Seattle, WA |
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Abstract: | Summary To provide a framework for biochemical investigation of ecdysteroid action inAedes albopictus mosquito cells, we examined the effect of 20-hydroxyecdysone on cell growth and morphology, synthesis of inducible proteins
(EIPs), and expression of a transfected gene regulated by a synthetic ecdysteroid response element. When cells were cultured
in the continuous presence of 10−6
M 20- hydroxyecdysone, the rate of growth decreased and subtle changes in cell morphology were observed. In bothAedes aegypti andA. albopictus cells, synthesis of a small number of radiolabeled proteins, which appeared as minor bands on sodium dodecyl sulfate-polyacrylamide
gels, was induced by treatment with 20-hydroxyecdysone. On two-dimensional polyacrylamide gels, 11 EIPs, ranging in size from
approximately 22 to 52 kDa, were identified inA. albopictus C7-10 cells. Ten inducible proteins were localized in the cytoplasmic fraction; EIP28 and EIP31 were detected in both cytoplasmic
and nuclear extracts, and EIP29 was detected only in the nucleus, at a very low level. None of these proteins corresponded
to small heat shock proteins, whose genes are 20-hydroxyecdysone-inducible in someDrosophila cell lines. The juvenile hormone analog, methoprene, induced expression of a 25 kDa protein in C7-10 cells. Although 20-hydroxyecdysone
sustained the synthesis of this methoprene-inducible protein, synthesis did not occur in the presence of 20-hydroxyecdysone
alone. In transfectedA. albopictus cells, expression of a recombinant DNA construct containing two tandem synthetic ecdysteroid regulatory elements based on
aD. melanogaster small heat shock protein gene was modestly induced by 20-hydroxyecdysone. |
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Keywords: | Aedes mosquito 20-hydroxyecdysone juvenile hormone |
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