Structure, expression and gene sequence of a juvenile hormone esterase-related protein from metamorphosing larvae of Trichoplusia ni.

Abstract

A carboxylesterase with an encoded molecular size of 61 kDa and a high sequence similarity to juvenile hormone esterase (JHE) has been cloned from cDNA prepared from final instar larvae of Trichoplusia ni. The absence of a recognizable encoded signal peptide suggests that the enzyme, JHER (for JHE-related) may not be secreted, in contrast to JHE. When the amino acid sequence of JHE, JHER and other esterases were mapped onto the secondary and tertiary structure determined crystallographically for acetylcholinesterase, certain structural features for the substrate binding/catalytic site were identified as common only to JHE and JHER. However, several differences between JHE and JHER were identified in residues at the binding/catalytic site, suggesting that although the two enzymes prefer similar natural substrates, these substrates are not identical. JHER is present as a single-copy gene, transcribed during the feeding stage of the final stage of the final larval stadium, but not after metamorphic commitment to the pupal developmental programme. The gene transcribes a single-size message of 2.0 kb. The genes for JHER and JHE appear to be physically juxtaposed in the T. ni genome. The 5' flanking sequence to the JHER gene possesses some sequences in common with the JHE gene, but is also missing some regulatory elements previously identified in the JHE gene. Sequences conserved between the promoters for the two genes were identified that were different from previously reported regulatory elements of eukaryotic transcription factors.