School of Life Sciences, University of Hyderabad, Hyderabad 500046, India.
Gen Comp Endocrinol. 2010 May 15;167(1):113-21. doi: 10.1016/j.ygcen.2010.02.007. Epub 2010 Feb 18.
The biologically active ecdysteroid hormone, 20-hydroxyecdysone (20E), regulates various processes like molting, metamorphosis and reproduction in insects. However, its role in expression of silk genes is obscure. The silk core in insects is generally constituted of a complex of three proteins namely, H-chain fibroin (H-fibroin), L-chain fibroin (L-fibroin) and P25. In the present study, we report full-length cDNA cloning and characterization of L-fibroin and P25 genes from rice moth, Corcyra cephalonica. Northern analyses demonstrated 1.1 and 1kb transcripts of L-fibroin and P25 respectively. The tissue expression pattern shows the presence of these transcripts specifically in the salivary gland. These two genes are developmentally regulated at transcriptional level and their maximum expression is observed during the late-last instar larval stage. Semi-quantitative and real-time PCR studies revealed that 20E regulates the expression of these genes in a dose-dependant manner. This study further shows that physiological dose of 20E does not alter the normal expression of these two genes, whereas treatments with higher doses cause a significant decline in the expression. This study clearly suggests the role of 20E in the regulation of L-fibroin and P25 at molecular level.
生物活性蜕皮激素 20-羟基蜕皮酮(20E)调节昆虫蜕皮、变态和繁殖等各种过程。然而,它在丝基因表达中的作用尚不清楚。昆虫的丝核心通常由三种蛋白质组成,即 H 链丝素(H-丝素)、L 链丝素(L-丝素)和 P25。本研究从米蛾 Corcyra cephalonica 中报告了 L-丝素和 P25 基因的全长 cDNA 克隆和特征。Northern 分析显示 L-丝素和 P25 分别有 1.1 和 1kb 的转录物。组织表达模式显示这些转录物特异性存在于唾液腺中。这两个基因在转录水平上受到发育调控,它们的最大表达发生在最后一龄幼虫晚期。半定量和实时 PCR 研究表明,20E 以剂量依赖的方式调节这些基因的表达。本研究进一步表明,生理剂量的 20E 不会改变这两个基因的正常表达,而高剂量处理会导致其表达显著下降。这项研究清楚地表明 20E 在 L-丝素和 P25 基因表达的分子水平上起调节作用。