Cloning and characterization of a new isoform of Choristoneura hormone receptor 3 from the spruce budworm

Abstract

Choristoneura hormone receptor 3 (CHR3) is a 20E (20-hydroxyecdysone)-induced delayed early gene that is homologous to Manduca hormone receptor 3 (MHR3), Drosophila hormone receptor 3 (DHR3), and Galleria hormone receptor 3 (GHR3). We recently cloned and characterized a cDNA that was copied from the 4.5 kb CHR3B mRNA. To isolate additional CHR3 isoforms, the Choristoneura fumiferana embryonic cDNA library was screened using CHR3B cDNA as a probe. Characterization and partial sequencing of 16 clones showed that one of them differed from the CHR3B in two regions. This cDNA (CHR3C) was completely sequenced; the sequence analysis showed that the longest open reading frame had 651 codons. The deduced amino acid sequence of this open reading frame contained all five domains that are typical for a steroid hormone nuclear receptor. The nucleotide sequence of CHR3C cDNA is identical to the nucleotide sequence of CHR3B cDNA except for two major differences in the A/B and D-domains. The CHR3C specific probes detected two mRNAs 5.4 kb (CHR3C), and 6.4 kb (CHR3D), which were present in the pupal stage. The CHR3C and CHR3D mRNAs are induced by the stable ecdysteroid analog RH-5992. The CHR3C protein also binds to the response element of the retinoic acid receptor-related orphan receptor.