Antisense expression of the 20-hydroxyecdysone receptor (EcR) in transfected mosquito cells uncovers a new EcR isoform that varies at the C-terminal end
G. Jayachandran et Am. Fallon, Antisense expression of the 20-hydroxyecdysone receptor (EcR) in transfected mosquito cells uncovers a new EcR isoform that varies at the C-terminal end, IN VITRO-AN, 37(8), 2001, pp. 522-529
The insect steroid hormone 20-hydroxyecdysone initiates a cascade of regula
tory events in a temporal and tissue-specific manner by first binding to a
complex of an ecdysone receptor (EcR) protein and a ultraspiracle protein.
Using an antisense (As) ribonucleic acid approach, we show that disruption
of EcR expression in transfected C7-10 cells from the mosquito Aedes albopi
ctus affects survival and growth. From stably transfected cells, we recover
ed a new isoform of A. albopictus AalEcRa, which is named AalEcRb. The dedu
ced amino acid sequence of AalEcRb was almost identical to that of AalEcRa,
with the exception of a seven amino acid sequence near the C-terminus. Usi
ng polymerase chain reaction followed by restriction enzyme analysis, we fo
und that AalEcRa is the predominant species expressed by wildtype C7-10 cel
ls, while cells transfected with As-EcR expressed both isoforms at approxim
ately equal levels.