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Cultured Aedes Albopictus Mosquito Cells Synthesize Hormone-Inducible Proteins

Que Lan, Anna Gerenday and Ann Marie Fallon
In Vitro Cellular & Developmental Biology. Animal
Vol. 29A, No. 10 (Oct., 1993), pp. 813-818
Stable URL: http://www.jstor.org/stable/4294134
Page Count: 6
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Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Cultured Aedes Albopictus Mosquito Cells Synthesize Hormone-Inducible Proteins
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Abstract

To provide a framework for biochemical investigation of ecdysteroid action in Aedes 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 both Aedes aegypti and A. 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 in A. 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 some Drosophila 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 transfected A. albopictus cells, expression of a recombinant DNA construct containing two tandem synthetic ecdysteroid regulatory elements based on a D. melanogaster small heat shock protein gene was modestly induced by 20-hydroxyecdysone.

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