Antisense expression of the 20-hydroxyecdysone receptor (EcR) in transfected mosquito cells uncovers a new EcR isoform that varies at the C-terminal end.
The insect steroid hormone 20-hydroxyecdysone initiates a cascade of regulatory 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 albopictus affects survival and growth. From stably transfected cells, we recovered a new isoform of A. albopictus AalEcRa, which is named AalEcRb. The deduced 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. Using polymerase chain reaction followed by restriction enzyme analysis, we found that AalEcRa is the predominant species expressed by wild-type C7-10 cells, while cells transfected with As-EcR expressed both isoforms at approximately equal levels.
Jayachandran
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Department of Entomology, University of Minnesota, St Paul, Minnesota 55108, USA. jayac001@tc.umn.edu
Fallon
A M
AM
D000330:Aedes / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D009033:Culicidae / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D016372:RNA, Antisense / Q000235:genetics*; D011987:Receptors, Steroid / Q000737:chemistry* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D020033:Protein Isoforms; D016372:RNA, Antisense; D011987:Receptors, Steroid; C034808:ecdysteroid receptor