Periodic expression of an ecdysteroid-induced nuclear receptor in a lepidopteran cell line (IAL-PID2).
A set of DNA primers was designed within the DNA-binding domain of the Manduca hormone receptor 3 (MHR3) cDNA. These primers were used in RT-PCR to isolate a 204 bp cDNA fragment from IAL-PID2 cells exposed to 10(-6) M 20-hydroxyecdysone (20E) for 12 h. The amino acid sequence deduced from the cDNA fragment presented 100% identity with the zinc finger domain of Manduca hormone receptor 3 (MHR3), Galleria hormone receptor 3 (GHR3) and Choristoneura hormone receptor 3 (CHR3). This cDNA fragment was used as a probe on total RNA from IAL-PID2 cells exposed to 20E and hybridized to mRNA, the size of which was close to 4.5 kb and named Plodia hormone receptor 3 (PHR3). Kinetics of induction of PHR3 mRNA were similar to that of HR3 genes but varied according to the position of cells in their cell cycle. The non-steroidal ecdysone agonist, RH-5992 induced the expression of PHR3 at lower concentrations than 20E. From sequence similarity, mRNA size, 20E and RH-5992 inducibilities, we conclude that PHR3 transcript could encode a Plodia hormone receptor 3 involved in the genetic signalling cascade of 20E. Thanks to its periodic expression, this putative orphan nuclear receptor could serve as a suitable cellular marker for studying changes of epidermal cell sensitivity to 20E during the cell cycle.
Debernard
S
S
Laboratoire de Physiologie Cellulaire des Invertébrés, Université Pierre et Marie Curie, 12 rue Cuvier, 75005, Paris, France. stephane.debernard@snv.jussieu.fr
Bozzolan
F
F
Duportets
L
L
Porcheron
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D001483:Base Sequence; D002453:Cell Cycle; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004441:Ecdysterone / Q000494:pharmacology*; D015870:Gene Expression; D006834:Hydrazines / Q000494:pharmacology; D007605:Juvenile Hormones / Q000494:pharmacology; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D011987:Receptors, Steroid / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013997:Time Factors
D018076:DNA, Complementary; D006834:Hydrazines; D007605:Juvenile Hormones; D012333:RNA, Messenger; D011987:Receptors, Steroid; D004441:Ecdysterone; D000841:Anisomycin; C082026:tebufenozide