The ultraspiracle gene of the Spruce Budworm, Choristoneura fumiferana: Cloning of cDNA and developmental expression of mRNA

Abstract

Cloning and characterization of a Choristoneura fumiferana ultraspiracle (Cfusp) cDNA are described. First, a PCR fragment and then a cDNA clone (4.4 kb) were isolated from spruce budworm cDNA libraries. Comparison of the deduced amino acid sequence of this cDNA with the sequences in Genbank showed that this sequence had high homology with the ultraspiracle cDNAs cloned from Drosophila melanogaster (Dmusp), Bombyx mori (Bmusp), Manduca sexta (Msusp), and Aedes aegypti (Aausp). The Cfusp cDNA contained all the regions that are typical for a steroid/thyroid hormone receptor superfamily member. The DNA binding domain or C region was the most conserved sequence among all the usps. The A/B, D, and E regions also showed high amino acid identity with the amino acid sequences of Dmusp, Msusp, Bmusp, and Aausp. The Cfusp 4.5-kb mRNA was present in the embryos, in all larval stages, and in the pupae. The Cfusp mRNA levels in the midgut increased during the sixth-instar larval development and reached peak levels during the ecdysteroid raises for the pupal molt. However, Cfusp mRNA levels remained unchanged in the midgut of fifth-instar larvae, and in the epidermis and fat body of sixth-instar larvae indicating both a tissue- and stage-specific regulation of Cfusp mRNA expression.

The sequence of Locusta RXR, homologous to Drosophila Ultraspiracle, and its evolutionary implications

“…Parsimony analysis of the protein sequences yielded one phylogenetic tree for the DBDs but a different tree (Oro et al 1990); Aae Aedes aegypti, (Kapitskaya et al 1996); Cte Chironomus tentans, AF045891; Bmo Bombyx mori, U06073 (Tzertzinis et al 1994); Cfu Choristoneura fumiferana, AF016368 (Perera et al 1998); Mse Manduca sexta, U44837 (Jindra et al 1997); Lmi Locusta migratoria; AF136372 (this study); Aam1 Amblyomma americanum RXR1, AF035577 (Guo et al 1998b); Aam2 Amblyomma americanum RXR2, AF035578 (Guo et al 1998b); Upu Uca pugilator (Chung et al 1998); Hsaa Homo sapiens RXRα, X52773 (Mangelsdorf et al 1990); Hsab Homo sapiens RXRβ, M84820 (Chambon et al 1992); Hsag Homo sapiens RXRγ, U38480 (Mangelsdorf et al 1992); Gga Gallus gallus RXRγ, X58997 (Rowe et al 1991); Xla Xenopus laevis RXRα, L11446 (Blumberg et al 1992); Dre Danio rerio RXRα, U29940 (Jones et al 1995). Numerals between abbreviations and sequences position of the first aligned amino acid in each protein.…”

“…1A. Homologs of Drosophila USP have now been cloned from a number of holometabolous insect species including two other dipterans, the mosquito Aedes aegypti (Kapitskaya et al 1996) and the midge Chironomus tentans (Genbank Accession AFO45891), and three moths, Bombyx mori (Tzertzinis et al 1994), Manduca sexta (Jindra et al 1997) and Choristoneura fumiferana (Perera et al 1998). Similar molecules are found in other arthropods including the tick, Amblyomma americanum (Arachnida: Ixodidae; Guo et al 1998b), and the crab, Uca pugilator (Crustacea: Decapoda; Chung et al 1998).…”

Cell cycle profiles of EcR, USP, HR3 and B cyclin mRNAs associated to 20E‐induced G2 arrest of Plodia interpunctella imaginal wing cells

“…Sequence comparison A high degree of amino acid identity with Choristoneura fumiferana USP (CfUSP) (Perera et al ., 1998), Manduca sexta USP (MsUSP) (Jindra et al ., 1997) and Bombyx mori USP (BmUSP) (Tzertzinis et al ., 1994) was observed in the C and E/F regions of PIUSP (Table 1).…”

Preparation of Functional Ecdysteroid Receptor Proteins (EcR and USP) Using a Wheat Germ Cell-Free Protein Synthesis System

“…These results clearly show that EcR and USP expressed with PRO-TEIOS bind to hsp27 EcRE as a heterodimer, as observed for EcR and USP of several insects prepared with rabbit reticulocyte lysate. 1,3,11,19,[35][36][37] Significant specific binding of PonA to EcR, but not to USP, was detected (Fig. 4).…”