Isoform specific control of gene activity in vivo by the Drosophila ecdysone receptor.
The steroid hormone 20-hydroxyecdysone induces metamorphosis in insects. The receptor for the hormone is the ecdysone receptor, a heterodimer of two nuclear receptors, EcR and USP. In Drosophila the EcR gene encodes 3 isoforms (EcR-A, EcR-B1 and EcR-B2) that vary in their N-terminal region but not in their DNA binding and ligand binding domains. The stage and tissue specific distribution of the isoforms during metamorphosis suggests distinct functions for the different isoforms. By over-expressing the three isoforms in animals we present results supporting this hypothesis. We tested for the ability of the different isoforms to rescue the lack of dendritic pruning that is characteristic of mutants lacking both EcR-B1 and EcR-B2. By expressing the different isoforms specifically in the affected neurons, we found that both EcR-B isoforms were able to rescue the neuronal defect cell autonomously, but that EcR-A was less effective. We also analyzed the effect of over-expressing the isoforms in a wild-type background. We determined a sensitive period when high levels of either EcR-B isoform were lethal, indicating that the low levels of EcR-B at this time are crucial to ensure normal development. Over-expressing EcR-A in contrast had no detrimental effect. However, high levels of EcR-A expressed in the posterior compartment suppressed puparial tanning, and resulted in down-regulation of some of the tested target genes in the posterior compartment of the wing disc. EcR-B1 or EcR-B2 over-expression had little or no effect.
Schubiger
Margrit
M
Department of Biology, University of Washington, Seattle WA 98195, USA. margrit@u.washington.edu
Tomita
Shuichiro
S
Sung
Carl
C
Robinow
Steven
S
Truman
James W
JW
D000818:Animals; D004330:Drosophila / Q000502:physiology*; D005786:Gene Expression Regulation; D017344:Genes, Insect; D008675:Metamorphosis, Biological; D009474:Neurons / Q000378:metabolism; D020033:Protein Isoforms / Q000502:physiology; D011987:Receptors, Steroid / Q000737:chemistry / Q000502:physiology*; D013997:Time Factors
D020033:Protein Isoforms; D011987:Receptors, Steroid; C055469:ecdysone receptor