| Popis: |
Juvenile hormone (JH) allows larval molting in response to ecdysteroids but prevents the switching of gene expression necessary for metamorphosis. I first review our efforts to isolate the nuclear receptor for JH in the larval epidermis of Manduca sexta using photoaffinity analogs and our recent findings that the molecule isolated does not bind JH I with high affinity. The reported apparent high affinity binding of JH I by the recombinant 29 kDa protein (rJP29) was artifactual due to the presence of contaminating esterases. Purified rJP29 bound little detectable JH I, but its binding of the photoaffinity analog was prevented by JH I as well as other isoprenoids, indicating a low affinity for these compounds. Our recent studies focus on the effects of JH on the early molecular events induced by 20-hydroxyecdysone (20E). Culture of day 2 5th larval epidermis with 10(-6)M 20E for 24 h caused first pupal commitment, then the onset of the predifferentiative events necessary for pupation. Biphasic increases in the mRNAs of the two isoforms of the ecdysone receptor (EcR-A and EcR-B1) and of E75A, an ecdysteroid-induced transcription factor, coincided with these two phases. The mRNAs for Ultraspiracle (USP) and the metamorphosis-specific Broad-Complex (BR-C) increased only during the second phase. The presence of JH had no effect on the initial increases in EcR mRNAs but caused an increased accumulation of E75A mRNA. This JH also prevented the later changes in EcR, USP, and BR-C mRNAs. Thus, JH influences only certain of the early actions of 20E which then result in its preservation of the "status quo." |
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