Identification and mRNA developmental profiles of two ultraspiracle isoforms in the epidermis and wings of Manduca sexta.

Abstract

cDNAs were isolated from Manduca sexta that encode two isoforms of an ultraspiracle (USP) homologue MsUSP-1 and MsUSP-2 with different N-terminal A/B regions. The MsUSP-1 cDNA predicts a protein with 97% and 45% amino acid identities in the DNA- and ligand-binding domains respectively to the Drosophila USP and 89% overall identity with Bombyx mori CF1 (an USP homologue). Northern blot hybridizations with probes specific to MsUSP-1 and MsUSP-2 showed transcripts of an approximately equal size (4.5 kb), but with diverse developmental profiles in Manduca epidermis during the two final larval instars and the onset of the adult moult. The MsUSP-1 mRNA was expressed during the intermoult periods, with higher levels around the time of the larval ecdyses and at the onset of wandering behaviour. In contrast, the MsUSP-2 mRNA was up-regulated at times of high ecdysteroid titre during the larval moults, when the MsUSP-1 mRNA disappeared. Together, these conversely regulated isoform mRNAs contribute to the constitutive expression profile of total MsUSP mRNA.