Debernard S, Bozzolan F, Duportets L, Porcheron P
Laboratoire de Physiologie Cellulaire des Invertébrés, Université Pierre et Marie Curie, 12 rue Cuvier, 75005, Paris, France.
Insect Biochem Mol Biol. 2001 Oct;31(11):1057-64. doi: 10.1016/s0965-1748(01)00053-4.
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.
在烟草天蛾激素受体3（MHR3）cDNA的DNA结合结构域内设计了一组DNA引物。这些引物用于逆转录聚合酶链反应（RT-PCR），以从暴露于10^(-6) M 20-羟基蜕皮激素（20E）12小时的IAL-PID2细胞中分离出一个204 bp的cDNA片段。从该cDNA片段推导的氨基酸序列与烟草天蛾激素受体3（MHR3）、粉斑螟激素受体3（GHR3）和云杉卷叶蛾激素受体3（CHR3）的锌指结构域具有100%的同一性。该cDNA片段用作探针，与暴露于20E的IAL-PID2细胞的总RNA杂交，并与大小接近4.5 kb的mRNA杂交，将其命名为印度谷螟激素受体3（PHR3）。PHR3 mRNA的诱导动力学与HR3基因相似，但根据细胞在细胞周期中的位置而有所不同。非甾体蜕皮激素激动剂RH-5992在比20E更低的浓度下诱导PHR3的表达。从序列相似性、mRNA大小、20E和RH-5992的诱导能力来看，我们得出结论，PHR3转录本可能编码一种参与20E遗传信号级联反应的印度谷螟激素受体3。由于其周期性表达，这种推定的孤儿核受体可作为研究细胞周期中表皮细胞对20E敏感性变化的合适细胞标记物。
